Targeted RNA-based therapeutics with multiple specificities and high efficacies are now available. In this review, we discuss progress in RNA-based antitumor therapeutics, including mRNAs, siRNAs, miRNAs, ASOs, saRNAs, RNA aptamers, and CRISPR-based gene editing. We focus on the immunogenicity, stability, translation efficiency, and delivery of RNA drugs, and summarize their optimization and the development of delivery systems. In addition, we describe the mechanisms by which RNA-based therapeutics induce antitumor responses. Furthermore, we review the merits and limitations of RNA cargos and their therapeutic potential for cancers.RNA-based therapeutics  have considerable potential for tumor treatment. The development and optimization of RNA modifications and delivery systems enable the stable and efficient delivery of RNA cargos  RNA-based therapeutics, including messenger RNAs (mRNAs), small interfering RNAs (siRNAs), microRNAs (miRNAs), antisense oligomers (ASOs), small activating RNAs (saRNAs), RNA aptamers, and CRISPR-based gene editing—have considerable therapeutic potential for genetic diseases These advantages are particularly beneficial in tumor treatment. Such therapeutics can control the expression of target proteins, reshape the suppressive tumor microenvironment (TME) by regulating cytokine expression, and induce an innate or adaptive immune response RNA-based therapeutics remains a significant challenge due to the inherent susceptibility of natural RNA to degradation by nucleases. As a result, there is a need to modify RNA structures and optimize delivery vehicles to efficiently deliver nucleic acid cargos to the target tissues and/or organs Here we review recent progress in antitumor RNA-based therapeutics—mRNAs, siRNAs, miRNAs, ASOs, saRNAs, RNA aptamers, and CRISPR-based gene editing. We focus on the mechanisms by which RNA-based therapeutics induce antitumor immune responses. We also discuss RNA modification strategies and classify their carriers according to composition. Moreover, we review the relative merits and bottlenecks of various RNA-based therapeutics in tumor treatment Figure .RNA molecules are inherently unstable. Exogenous RNA molecules trigger immune responses, leading to limited level of protein expression. RNA molecule modifications can be used to address these issues, which is the subject of the next section.in vitro transcription (IVT). They consist of five main structures: the 5' cap, the 5' and 3' untranslated regions (UTRs), the open reading frames (ORFs) encoding the target proteins, and the 3' poly(A) tail. These structures affect the stability, translation efficiency, and immunogenicity of mRNA-based therapeutics mRNA drugs are synthetic versions of mature eukaryotic mRNAs and are typically produced by The 5' and 3' UTRs are non-coding regions whose secondary structures, elements, and lengths affect ribosome recruitment and mRNA translation The 5' cap structure contributes to mRNA stability and translation efficiency in vivo. Activation of Toll-like receptor (TLR) 3 can be inhibited by replacing the original nucleotide with 6-methyladenosine (m6A) or 2-thiouridine (s2U), whereas replacement with 5-methylcytidine (m5C), 5-methyluridine (m5U), s2U, m6A, or pseudouridine (Ψ) blocks the activation of TLR7 and TLR8, thus preventing an innate immune response 5C enhance RNA stability and translational capability while diminishing its immunogenicity 1-methyl Ψ-modified nucleotides are employed in IVT to improve the safety and stability of mRNA vaccines. Two mRNA vaccines against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) were modified using N1-methyl Ψ and each showed > 90% protection against coronavirus disease 2019 (COVID-19) Nucleotide modification can also optimize mRNA stability, translation efficiency, and immunogenicity siRNA can be chemically modified to enhance the selectivity of their antisense strands for RNA-induced silencing complex (RISC) loading and to reduce off-target RNAi activity. However, unmodified siRNA may induce immune toxicity by activating TLR3. A variety of chemical modification strategies can be used to maximize the therapeutic efficacy and minimize the side effects of siRNAs.Chemical modification typically targets the sugar ring, base, and phosphate skeleton of nucleotides. The goals are to improve the binding affinity of siRNA and protect it from nuclease degradation The most common phosphate skeleton modification is phosphorothioate (PS), which prolongs the half-life in the circulation. Clinically approved siRNA therapeutics  have PS modifications. The phosphorodithioate (PS2) substitution in siRNAs involves the replacement of two non-bridged oxygen atoms. The potency and nuclease resistance of PS2-modified siRNAs are slightly higher than those of PS and unmodified siRNAs. Boron phosphate siRNA may be more effective than phosphorothioate 5C and m6A, tend to be used Base replacement is another modification strategy for siRNA-based therapeutics. 5'-nitroindole modification of siRNAs reduces passenger strand-mediated off-target effects N-acetylgalactosamine -siRNA conjugates enable drug delivery to the liver. Tris-GalNAc binds to asialoglycoprotein receptor, which is highly expressed on the surface of hepatocytes, resulting in rapid endocytosis ASOs are susceptible to nuclease degradation An example of a modified ASO molecule is peptide nucleic acids (PNAs) In vivo administration of 5'-cholesterol-modified aptamers significantly prolongs plasma half-life and exposure The inherent susceptibility of RNA aptamers to nuclease degradation determines their stability 2, 2'-LNA Sp or Rp configuration), unlike the natural conformation, and may therefore have adverse effects on biological function.Modifications of the sugar ring can improve nuclease stability and prolong serum half-life. Similarly, the 2'-OH positions of RNA aptamers can be substituted with 2'-F m +1.6°C, higher than the 2'-OMe modification) in vivom by mediating hydrophobic stacking interactions Chemical modification strategies are under development for other RNA-based therapeutics. The nucleotide sugar ring, base, and phosphate skeleton are modified in anti-miRNA oligonucleotides (AMOs). Most widely used chemical modifications on AMOs are LNA, 2'-F-RNA, 2'-OMe, PNAs. Modified AMOs with higher binding affinity and superior stability have greater regulatory potency RNA therapeutics must be delivered to the correct tissues without triggering an immune reaction. However, the high molecular weight and negative charge of RNA hampers their delivery to target sites. RNA delivery systems can be divided to viral or nonviral. Below we summarize several nonviral RNA delivery systems being extensively studied Figure .in vivo transfection capability and reduce their toxicity Polymeric NPs are typically prepared from biocompatible and biodegradable polymers, in which the drug is dissolved, entrapped, encapsulated, or attached to a nanoparticle matrix. Polymers bind nucleic acids to form polymeric complexes at physiological pH, facilitating gene delivery. Polymeric NPs promote electrostatic binding to nucleic acid cargo by interacting with positively charged units. Nucleic acids and polymers can be covalently linked using degradable linkers. The addition of cationic groups such as chitosan to polylactic-co-glycolic acid (PLGA) enables its use for siRNA delivery Lipid-based NPs have high biocompatibility and biosafety, and their production is simple. They include liposomes, micelles, emulsions, and solid lipid NPs (SLNs). LNPs have been evaluated as mainstream delivery systems in multiple preclinical trials.Because the size, shape, surface charge, and materials of NPs affect their cellular uptake Inorganic NPs  have a narrow size distribution and a surface chemistry suitable for ligand conjugation in vivo for extending the plasma half-lives of AuNPs and enhance their targeted accumulation and controllable release AuNPs typically have diameters of 1 to 100 nm, a large surface-to-volume ratio, good optoelectronic properties, excellent biocompatibility, and low toxicity MSNs are biodegradable and chemically stable nanostructured materials composed of silica particles with pore channels Versatile multifunctional nanomaterials whose synthesis is simple and inexpensive, as well as hybrids that integrate the advantages of different materials, have been developed in vivo delivery of RNA-based therapeutics in clinical applications.Hydrogels consist of a three-dimensional network swollen with water in vitro production and purification of mRNAs prevents host protein and virus contamination in vivo degradation and promote the intracellular delivery and endosomal escape of mRNAs mRNA-based therapeutics are promising alternatives to DNA for cancer immunotherapy due to their lower mutagenicity and easier transient expression. Furthermore, the LNPs consist of phospholipids, cholesterol, PEGylated lipids, and cationic or ionizable lipids. Phospholipids and cholesterol mediate LNP endocytosis and accelerate mRNA release during endocytosis. PEGylated lipids prolong the half-life in the circulation Acute inflammatory responses, such as pain, swelling, and fever, can be caused by mRNA-LNP vaccines  et al.in vitro, enhancing CD11c-cell maturation and CD8 T-cell proliferation in aggressive E·G7-OVA lymphoma models Depending on their charge and composition, LNPs can be broadly classified as cationic LNPs, ionizable LNPs, and lipid calcium phosphate (LCP) NPs. Classical mRNA cancer vaccines target tumor-associated antigens (TAAs) preferentially expressed in malignant cells. For instance, Oberli et al.in vivoSeveral ionizable lipoplex-type mRNA carriers are available. Tateshita et al. et al. reported that a decreased cationic lipid-to-DOPE ratio of mRNA-loaded lipoplexes affected organ specificity. Based on this rationale, they developed a lipoplex-based system for cargo delivery to splenic DCs The clinical applications of the above delivery system are hampered by the lack of specific targeting, which make them useful only for local inoculation and liver-targeted therapy. Efficient lymphatic drainage and accumulation can be promoted by PEGylation and modifying NP size and surface charge Use of mRNAs to restore tumor-suppressor expression has therapeutic potential for cancer. An example is a polymeric NP platform for delivering mRNA encoding phosphatase and tensin homolog deleted on chromosome ten (PTEN), a cancer-inhibiting factor. Reactivating PTEN in PTEN-mutated melanoma cells and PTEN-null prostate cancer cells by mRNA delivery reversed the immunosuppressive TME by promoting CD8 T-cell infiltration and lifting the expression level of proinflammatory cytokines in vivo. The CXCR4-targeting NP system transported p53 mRNA to HCC cells, restoring p53 activity and decreasing HCC cell viability. This mRNA nanotherapy-based p53 restoration strategy in combination with anti-PD-1 therapy induced a potent antitumor effect in intrahepatic and ectopic models of HCC with p53 loss. Therefore, combining p53 mRNA therapeutics with ICB could reverse immunosuppression in HCC p53 is a tumor suppressor involved in cell cycle arrest, apoptosis, senescence, and other cellular pathways + TiDCs in vivo into stimulatory cells that efficiently process spontaneously engulfed TAAs, upregulate costimulatory molecules, and migrate to TDLNs to activate cytotoxic T lymphocytes (CTLs), ultimately delaying the growth of established tumors mRNAs encoding cytokines or chemokines can also induce APC maturation and activation, activate T-cell-mediated immunity, and adjust the dysfunctional immune TME. The mRNA-based adjuvant TriMix consists of mRNAs encoding the costimulatory molecule CD70, the activation stimulator CD40 ligand (CD40L), and constitutively active TLR4 (caTLR4). Upon codelivery of tumor-associated antigen (TAA), the TriMix mRNA reprograms CD8+ T-cell infiltration and IFN-γ expression. The systematic administration of such cytokines leads to high exposure, possibly resulting in toxicity. For this reason, MEDI1191, an optimized IL-12 mRNA, was developed for tumor-targeted local delivery. To minimize off-target liver toxicity, the miRNA-mediated binding site was incorporated into the 3'-UTR of the IL-12 mRNA to promote its elimination Moderna has collaborated with AstraZeneca to develop a local intratumoral mRNA therapy for IL-12 delivery γ mRNAs and is intended for the treatment of lymphoma (NCT03739931). OX40L enhances the expansion and survival of CD4 and CD8 T cells, and IL-23 and IL-36γ are pro-inflammatory cytokines of the IL-12 and IL-1 families, respectively, which activate and mature DCs and other immune cells. Compared to mono-cytokine mRNA therapy, addition of an mRNA encoding the T-cell costimulator OX40L increased the complete response rates of treated and untreated distal tumors. Mice treated with the mixture exhibited complete immune responses and effective protection Preclinical data suggest limited antitumor activity for IL-12 mRNA monotherapy. For this reason, other mRNA therapeutics encoding mixtures of cytokines and chemokines have been developed. Intratumoral injection of DAL4-LNP-IL-12 and IL-27 mRNAs showed synergistic suppression of tumor growth with robust infiltration of immune effector cells Engineered T cells that express chimeric antigen receptors (CARs) for adoptive cell therapy (ACT) have considerable benefits for the treatment of certain blood malignancies Clinical-scale manufacturing of engineered T cells requires their isolation, transfection, modification, amplification, and re-injection, which is difficult and costly. Although virus-mediated transfer can prolong transgene expression by T cells Nanocarrier-mediated targeted delivery of an mRNA encoding a rare-cleaving megaTAL nuclease disrupts T-cell receptor expression ex vivoIonizable lipid NPs have been used to deliver mRNAs to primary human T cells in vivo delivery efficiency.Most solid malignancies failed to effectively respond to CAR-T cell infusion because of tumor resistance, tumor-antigen-escape relapse, and the suppressive TME In one study, a liposomal antigen-encoding RNA was intravenously administered to stimulate tumor-associated T cells in patients with cancer CAR targeting macrophages rather than T cells has emerged as another meaningful strategy for the treatment of solid tumors. MT-101 is a new class of non-T cell CARs produced by transforming monocytes with mRNAs. Most monocytes in blood differentiate into macrophages after migrating to tissues. MT-101 targets tumor cells in peripheral tissues by expressing CARs targeting CD5 on the surface of monocytes. A phase 1/2 clinical study demonstrated the safety and tolerability of MT-101 in patients with refractory or relapsed peripheral T-cell lymphoma (PTCL) at day 28, with no dose-limiting toxicity, cytokine release syndrome (CRS), or immune effector cell-associated neurotoxicity syndrome (ICANS) (NCT05138458). In 2020, Moderna and Carisma Therapies established a cooperative relationship to combine Carisma's engineered macrophage technology with Moderna's mRNA and LNP technology and launched the first clinical study of their so-called CAR-M therapy (CT-0508). The combination of CT-0508 with an anti-PD-1 antibody (pembrolizumab) was used in the phase 1 clinical development stage to treat solid tumors with HER-2 overexpression.siRNAs are double-stranded RNA molecules of 21 to 23 nucleotides, typically with two free bases at the 3'-end, that silence target genes by RNA interference. The precursor is recognized by Dicer RNase and subsequently binds to the target mRNA via the RISC and cleaves it at bases 10 to 11 from the 5'-end, resulting in post-transcriptional gene silencing  et al.2O3 and HER2-siRNA shows an excellent antitumor effect in an orthotopic gastric tumor model. As2O3 induces apoptosis and suppresses tumor metastasis, and HER2-siRNA blocks the expression of the oncogene HER2, inhibiting tumor invasion and metastasis siRNAs can be used in combination with nanomaterials. Currently, various lipid-based delivery systems have been used for co-delivering siRNA and drugs Table . CationiSuitable modification of nanocarriers enables siRNA delivery to target tissues for cancer treatment. One study introduced a versatile codelivery platform for the treatment of triple-negative breast cancer  et al.Silencing key elements of tumor progression or downregulating immunosuppressive genes can induce an antitumor immune response  et al.+ T-cell activation, enhancing the effectiveness of cancer immunotherapy and inhibiting HCC progression in situ-injectable chitosan hydrogel containing C-C chemokine ligand 5 (CCL5) siRNA-loaded NPs, together with mRNA encoding lipid-immune regulatory factor 5 (IRF5), reshaped the TME in a model of pancreatic cancer by promoting M1 macrophage polarization Research is now focusing on combination therapy or codelivery via nanomaterials with siRNA. Guo et al.in vitro and in vivo. Coadministration of nanocarriers loaded with Xkr8 siRNA and the FuOXP prodrug into tumors via intravenous injection significantly inhibited tumor growth in colon and pancreatic cancer models and enhanced antitumor immune responses. In addition, doxorubicin (Dox) induces ICD in tumors, a type of apoptosis that enhances the protective immune response. Stable nucleic acid-lipid particles (SNALPs) have been used for the simultaneous delivery of Dox and an siRNA that knocks down CD47 (siCD47) Combination therapy with siRNA and chemotherapeutic drugs has shown promise for some types of cancer. ChenASOs are synthetic single-strand nucleic acids of 12 to 30 nucleotides. Upon entering a cell, they specifically bind to the target mRNA, forming a DNA-RNA hybrid that triggers mRNA cleavage via recruitment of RNase H, reducing the mRNA level  et al. used a 2.5-generation cEt-ASO (AZD9150) to suppress STAT3 expression, resulting in antitumor activity in lymphoma and lung cancer models Because conventional methods cannot downregulate the protein level of STAT3, Hong et al.The limited cell membrane permeability and lack of nuclear targeting of ASOs hinder their clinical application. To address these issues, ChengmiRNAs are single-stranded, non-coding RNAs typically of 20 to 24 nucleotides. They complementarily bind the 3' UTR of the target mRNA, and then cleave the mRNA or inhibit its translation in vitro and in mouse models of cancer. In addition, mice treated with MRX34 exhibited significant tumor growth inhibition, and a significantly increased survival rate. However, liposomal delivery systems have issues with liver and kidney accumulation and acute hypersensitivity reactions.The first clinical miRNA-based therapeutic was the liposomal formulation of miR-34a, named MRX34 Inorganic NPs are superior to liposomes in terms of their adjustable size and superior pharmacokinetics. For example, a lipid-coated calcium phosphonate NP has been designed for macrophage-targeted miRNA delivery in tumor immunotherapy Vesicular exosomes could be used for miRNA delivery. Exosomes, the smallest extracellular vesicles (EVs) (40 to 160 nm), can transport miRNAs in a paracrine and endocrine manner Small activating RNAs (saRNAs) are small dsRNAs of approximately 21 nucleotides. After generating active Ago-RNA complexes, they trigger gene expression activation by targeting promoter regions. In addition, they have an effect in any genomic region with antisense transcripts Lipid-based delivery systems are commonly used for systemic or local saRNA delivery. Take target gene P21 as example, results illustrated that using targeting saRNA transcriptionally activated p21 expression and promoted cell-cycle arrest in the G1/G0 phase, inhibiting cell proliferation and enhancing chemosensitivity Surface modification, for example, with biodegradable polymers such as PEG, can overcome the toxicity and off-target effects of LNP delivery systems RNA aptamers are sequences 20-100 nucleotides in length with complex three-dimensional structures that bind to target molecules with high affinity and specificity via non-covalent pocket interactions Several clinical trials of aptamers for tumor therapy are ongoing. The guanosine-rich AS1411 is the first oligodeoxynucleotide aptamer to be evaluated in phase I and II clinical trials. It suppresses tumor-cell proliferation by interfering with the stability of Bcl-2 mRNA RNA aptamers are used in combination with other therapeutic agents  for targeted delivery. For example, the combination of a PSMA-specific aptamer for prostate cancer with therapeutic oligonucleotides inhibited oncogene activity in PSMA-expressing cells, thereby having an antitumor effect The clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system has unprecedented therapeutic potential for genetic diseases The two critical components of the CRISPR/Cas9 system are the Cas9 protein and gRNA. The principle of this system involves the insertion of a specific DNA sequence (spacer) from the invading virus and plasmid into the CRISPR locus. Upon re-infection, the CRISPR sites containing the spacer acquired previously are transcribed and the products are processed into mature gRNAs. The gRNA targets Cas9 to a particular genomic locus, where it induces double-strand breaks (DSBs). For binding to and cleavage of DNA by Cas9, the 3'-side of the target sequence must have an NGG protospacer adjacent motif (PAM)  et al.Cancer arises from the accumulation of genetic/epigenetic abnormalities. CRISPR-Cas9-mediated genome editing enables precise manipulation of a genomic sequence, enabling the identification of genes involved in carcinogenesis and correction of oncogenic mutations  et al.in vivo, promoting tumor-cell apoptosis. In addition, the technique allows antibody-targeted delivery for the treatment of diffuse tumors. Intraperitoneal injection of EGFR-targeted sgPLK1-cLNPs resulted in selective uptake into diffuse ovarian tumors, leading to up to ~80% gene editing in vivo, inhibiting tumor growth and improving survival  et al.However, the CRISPR/Cas9 system has limitations related to cell injury, limited packaging capacity, and immune activation. In addition, the large sizes of Cas9  and sgRNA  preclude the use of viral and nonviral delivery systems in vivo delivery system have been developed, RNA-based drugs can be rapidly designed and synthesized. However, several technical bottlenecks affect RNA-based antitumor therapies, including their targeting specificity, safety, and efficacy. To address these issues, a variety of optimization and modification strategies for RNA molecules and delivery systems have been explored.Although cell and antibody-based therapies currently dominate tumor immunotherapy, more work is needed to overcome the risk for side effects, induction of an inappropriate and potentially harmful immune response, the complexity of treatment, and the high cost of production. RNA-based therapeutics  can modulate the expression of target genes to varying degrees. In addition, they can stimulate an immune response or reshape the suppressive TME by producing antigens or restoring the levels of beneficial proteins. The ability to target multiple genetic components is an advantage of RNA therapeutics over other small-molecule or protein-based drugs. Furthermore, once the chemical structure of the RNA molecule and the Therapeutic effectiveness is influenced by off-target effects induced by nonspecific accumulation. To prolong accumulation at tumor sites, promote target uptake, and control drug release, surface modifications with targeting ligands have been developed The safety of RNA cargos is a key consideration for their clinical application. The innate immunostimulatory properties of RNA molecules and their function as immune adjuvants enhance the immune response to antigens during vaccine development. However, excessive immunogenicity can lead to severe adverse reactions. Clinical translation requires the striking of a balance between safety and efficacy. The development of more biocompatible delivery vehicles with better biodegradability would accelerate the clinical translation of nanomedicines in vivo targeting of immune cells. Such approaches could enhance the effectiveness of immunotherapy and so reduce the RNA-based drug dose required. Because drugs have different pharmacokinetic and pharmacological characteristics, as well as interindividual differences in drug distribution after systemic administration, multiple-drug treatment strategies must consider the optimal dosage ratio and sequential release to reduce toxicity and enhance efficacy. P-gp inhibitors should be released by delivery systems prior to any co-delivered drugs. Otherwise, the co-delivered drug(s) might be exported extracellularly by P-gp, thus reducing their therapeutic efficacy.Another crucial issue is therapeutic effectiveness. Tumorigenesis involves complex regulatory networks and immune-escape mechanisms. Treating tumors with a single modality is often unsatisfactory. Treatment of tumors requires multiple modalities, potentially including codelivery of cytokines and chemokines, alleviating immunosuppressive signaling in the TME, and In conclusion, we reviewed the modifications of, delivery systems for, and potential applications of RNA-based therapeutics for cancers. Although RNA delivery platforms have limitations, advances in RNA-based bioengineering mean considerable therapeutic potential for cancer treatment. Indeed, these modalities offer hope for types of cancers with few or no treatment options.


Severe cutaneous adverse reactions (SCARs) including acute generalized exanthematous pustulosis (AGEP) are usually medication-related and represent a rare, heterogeneous group of T-cell-triggered delayed hypersensitivity reactions resulting from interactions between small molecule drugs, human leucocyte antigen (HLA) Class I molecules, and T-cell receptors (TCR) –5.18F]FDG PET/CT scan, performed four days after initial symptoms, demonstrated a very striking pattern of uniformly increased metabolic uptake throughout the skin. A marked [18F]FDG uptake was observed in the spine and spleen  due to systemic inflammatory response. Moreover, the neck and groin areas exhibited also an intense tracer uptake  mainly due to a partial volume effect and, to a lesser extent, physical stress in the skinfold areas in addition to the abovementioned dermatological changes. The elevated [18F]FDG uptake at the injection site in the right wrist is also to be mentioned. Skin biopsy revealed a superficial perivascular dermatitis with infiltration of the outermost layer of the dermis by lymphocytes, histiocytes, and single plasma cells, accompanied by focal parakeratosis and minimal spongiotic loosening of the stratum corneum (e).A 62-year-old male patient with an anaplastic large T-cell lymphoma in medical history presented in our clinic with an abrupt onset of generalized maculopapular exanthema without mucosal involvement after initiating antibiotic therapy with a combination of caspofungin, vancomycin, and meropenem due to community-acquired pneumonia. A whole-body [18F]FDG PET/CT scan.To our best knowledge, this is the first metabolic imaging of SCAR syndrome, in particular meropenem-induced AGEP, using [


On 5 May 2023, after more than 3 years of battle against COVID-19, the World Health Organization (WHO) officially announced the end of the global Public Health Emergency for COVID-19 . HoweverOur Research Topic addresses two emerging challenges encountered by COVID-19 patients: the re-detectable positive (RP) phenomenon and the long-term effects commonly referred to as long COVID. Furthermore, first-time population-based pharmacovigilance analysis for autoimmune hepatitis following COVID-19 vaccination was reported, and prospective therapeutic targets with a focus on PI3K/Akt/Nrf2 mediated cellular signaling pathway were discussed. We aim to provide valuable insights into the continuing challenges arising from this global pandemic.Peng et al. studied SARS-CoV-2 recurrence in individuals who have previously recovered from COVID-19 infection. Blood cell transcriptome sequencing reveals slight genomic discrepancies. In this study, the most distinguishing factors between re-detectable positive (RP) and non-re-detectable positive (NRP) patients were sex, IgG and IgM titers in the blood. The number of female RP patients was substantially high (67.8%), and the titers of IgG and IgM antibodies were also significantly raised. Differentially expressed genes (DEGs) were mainly involved in antigen recognition, inflammatory modulation, and immune responses. Co-expression networks from two mutually verified RP datasets were constructed, and 59 overlapping indicator genes were highly correlated with RP results. Therefore, along with long-term isolation and multiple nucleic acid tests, transcriptional sequencing can be a useful complementary strategy for the early and reliable diagnosis of RP patients.Kenny et al. highlighted the multifaceted nature of long COVID, encompassing respiratory, cardiovascular, neurological, and immunological manifestations. A thorough investigation is conducted in this all-encompassing review on the clinical characteristics, potential pathogenesis, and therapeutic targets for long COVID and they suggested pathogenic pathways involve immune dysfunction, sustained viral reservoirs, and tissue damage. Individuals with long COVID-19 have been observed with persistent T cell activation and alterations in immune populations, such as a decrease in the number of naïve B cells. Increased innate cell activation, altered cytokine levels, tissue viral persistence, autoimmunity, dysbiosis, dysautonomia, and endothelial dysfunction have also been linked to the development of long COVID-19.Chen et al. reported a total of 53 cases of autoimmune hepatitis (AIH) which were identified between 11 December 2020 and 15 March 2022 collected from the Centers for Disease Control (CDC) COVID Data Tracker and the Vaccines Adverse Event Reporting System (VAERS) after administration of COVID-19 vaccine. The overall rate of AIH that was reported in relation to COVID-19 vaccination was 0.21 (95% CI 0.16–0.27) per million individuals. The findings reveal that the AIH cases were lower than expected suggesting that post-vaccination autoimmune hepatitis cases may not pose a safety concern for COVID-19 vaccination.Prompt reporting of potential adverse events and implementation of robust pharmacovigilance systems are critical in the ongoing global vaccination campaigns. The administration of COVID-19 vaccinations has led to a rapid emergence of anecdotal reports on autoimmune hepatitis (AIH) following vaccination . ReportsLekshmi et al. reviewed the molecular mechanisms underlying the SARS-CoV-2 infection process, identifying potential therapeutic targets associated with the PI3K/Akt/Nrf2 cellular signaling pathway. The activation of Nrf2 has been demonstrated to alleviate the inflammatory response and oxidative stress induced by SARS-CoV-2 infection. The modulation of this pathway holds significant potential for the development of effective therapeutic interventions against COVID-19.In summary, this Research Topic offer valuable insights into various aspects of COVID-19 such as recurrence, long COVID, vaccine-related adverse effects and potential therapeutic targets. These studies significantly contribute to the existing knowledge base as the global scientific community continues to deepen its understanding of this virus. Translating these research findings into clinical practice empowers healthcare professionals to optimize strategies for managing COVID-19 and its long-term implications. Hence, continuous research efforts remain crucial in our collective fight against this unprecedented global health crisis.


Coronavirus disease 2019 (COVID-19) is a respiratory illness caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus that can induce myopathy, which can evolve into potentially life-threatening muscle weakness, including diaphragmatic paralysis. We present a case report of a 57-year-old female treated in the medical ICU for acute respiratory distress syndrome (ARDS) triggered by active COVID-19 infection, who subsequently developed worsening respiratory weakness from underlying COVID-19 myopathy manifesting as respiratory muscle weakness. Our patient’s muscle biopsy highlights the development of muscle atrophy without evidence of inflammatory myopathy, making the presence of pre-existing autoimmune myopathy unlikely. While literature cites different biochemical etiologies for the development of myopathy, the exact mechanism behind this phenomenon is not yet defined. Coronavirus disease 2019 (COVID-19) is a respiratory illness caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus that has been shown to induce myopathy, which can evolve into potentially life-threatening muscle weakness, including diaphragmatic paralysis . While lOur patient was a 57-year-old female with a past medical history of hypertension and obesity, who came to the emergency room with complaints of shortness of breath and chest tightness during the high COVID-19 pandemic. She described having these symptoms for the past week, with a gradual worsening in dyspnea and generalized muscle weakness leading to her visit. She denied chest pain, recent immobilization, surgery, travel, contraception use, or any history of cancers, deep vein thrombosis, or previous pulmonary emboli. She had no smoking history or any pertinent family history.Upon examination in the emergency room, she was hypoxic, with a resting room air saturation of 90% with moderate work of breathing, which subsequently improved to 95% after placing her on 2 liters of the nasal cannula. She was otherwise normotensive and afebrile. The patient's initial neurological exam revealed no cranial nerve deficits; however, she had generalized weakness in both upper and lower extremities, with strength graded at 3/5 in the upper extremities and 4/5 in the lower extremities. Her sensation was intact in all limbs. Her labs were significant for acute kidney injury, denoted by an elevated creatinine of 3.2 (baseline 0.9), along with high inflammatory markers low-density lipoprotein at 486 (standard 100-190 units/L), C-reactive protein at 93 (standard <5mg/L), D-dimer at 295 ng/mL (standard 0-230 ng/mL), and ferritin at 770 (standard 13-150 ng/mL). Initial chest x-ray revealed interstitial opacifications in the perihilar and lower lobe regions. The patient was admitted to the general medical unit as a person under investigation for COVID-19 and for the management of hypoxia in the setting of suspected community-acquired pneumonia. Her COVID-19 polymerase chain reaction tests came back positive.In the ward, she was started on empiric antibiotic therapy and hydroxychloroquine therapy of 400 mg daily. She continued to experience worsening hypoxia in the following days with respiratory distress and tachypnea, leading to a need for mechanical respiratory support. She received tocilizumab and was managed for acute respiratory distress syndrome (ARDS). She was successfully extubated to non-invasive positive pressure ventilation (NIPPV) after 48 hours of mechanical ventilation but showed signs of respiratory muscle weakness, with incentive spirometry showing reduced lung volumes below 500mL suggestive of diaphragmatic weakness. Her hospital course was complicated by severe pulmonary edema and the development of acute heart failure, suspected to be secondary to COVID-19 myocarditis. Despite intravenous diuretics and preload and afterload reduction, there was no improvement in the patient’s fraction of inspired oxygen (FiO2) requirements or radiologic studies. She received convalescent plasma and completed an entire course of the antibiotic regimen. However, she continued to exhibit respiratory muscle weakness and rapid tiring off NIPPV, with subsequent development of hypoxia and tachypnea. Repeat chest x-rays showed interval worsening of bilateral pulmonary infiltrates with reduced lung volumes despite diuresis and completion of the antibiotic regimen. Given these findings, the patient was re-intubated.Blood gas analysis at the time showed an elevated carbon dioxide level of 93 mmHg (standard 35-45 mmHg) and pH of 7.15 . Her oxygen requirements have also increased dramatically to 80% oxygen content on mechanical ventilation, with a congruent decline in hemodynamic stability, leading to the necessity of pressors and fluid resuscitation in the shock setting. The patient’s persistent hypoxia, elevated work of breathing, and declining respiratory status in the ICU were believed to be associated with her progressive diaphragmatic weakness, which led the ICU team to send myositis marker work-up along with a muscle biopsy to evaluate for possible underlying autoimmune-myositis. The patent’s laboratory studies showed negative anti-mitochondrial antibodies, anti-Jo-1 antibodies, smooth muscle antibodies, and DNA antibodies. Creatine kinase levels peaked at 330 units/liter (standard 20-200 units/liter), and no severe persistent electrolyte abnormalities were noted on her chemistry panels. Muscle biopsies were taken from the patient’s left quadriceps muscle. While being managed for her ARDS, our patient suffered a cardiac arrest which led to her passing despite resuscitative efforts.Figures COVID-19-induced myopathy is just one form of a broader spectrum of inflammatory muscle diseases. Other etiologies, such as idiopathic inflammatory myopathy and those that stem from endocrine disturbances and toxic substances, can also present with sudden onset progressive muscle weakness. In the case of inflammatory myopathies, a triggered immunological reaction results in muscular weakening and inflammation . CertainViral myositis occurs exclusively in the setting of viral infections, with common etiologies being enteroviruses, cytomegalovirus (CMV), Epstein-Barr virus (EBV), and human immunodeficiency virus (HIV) . TypicalCOVID-19-associated myopathy is present in up to 50% of hospitalized patients with the virus -21. It iOur patient’s muscle biopsy highlights the development of muscle atrophy without evidence of inflammatory myopathy, making the presence of pre-existing autoimmune myopathy unlikely. Furthermore, the presence of musculoskeletal weakness even before admission to the hospital and prolonged ICU course make CIM and CIP the less likely primary pathology. CIM presents, however, with the loss of thick filaments , while CCOVID-19-induced myopathy is a rare but significant complication that should be considered in patients presenting with muscle weakness or wasting, especially those with a history of COVID-19 infection. With its long list of differential diagnoses in infected patients, diagnosing without direct muscle biopsy visualization can be challenging. There are no specific therapies targeting COVID-19-induced myopathy at this time, yet due to its high prevalence, further research on this topic can help physicians better understand its pathogenesis. This may lead us closer to developing effective treatment to prevent potential long-term neuromuscular sequala of the disease.


The worldwide spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in early 2020 led to the coronavirus disease 2019 (COVID-19) pandemic. Acute lung diseases, such as COVID-19 pneumonia, can trigger stress cardiomyopathy, raising concerns about potential cardiovascular complications related to these diseases. The current case involved a 72-year-old man with SARS-CoV-2 infection who was experiencing dyspnea, desaturation, and oppressive retrosternal chest pain. On his admission to the hospital, an electrocardiogram demonstrated sinus tachycardia, negative T waves in leads V4-V6, and slight ST-segment elevation in the same precordial leads. The patient also had an increased troponin I value and worsening of his baseline respiratory failure, which required starting noninvasive ventilation. The echocardiogram showed moderately depressed left ventricular systolic function and apical ballooning. The echocardiographic changes resolved during hospitalization without directed therapeutic intervention. We diagnosed Takotsubo syndrome associated with SARS-CoV-2 infection; however, the pathophysiological disruption remains to be clarified. Following the description of the first cases in China in late 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection spread worldwide in just a few months, instigating fear and anxiety and raising concern about health complications such as cardiovascular disease risk . AdditioA 72-year-old man presented to the emergency department (ED) for acute-onset chest pain and dyspnea. At admission, the patient reported feeling extremely anxious because some of his relatives had tested positive for SARS-CoV-2 infection. During the night before his admission, the patient noticed desaturation on a portable oximeter that was resistant to higher oxygen flows. The patient grew increasingly dyspneic and anxious owing to his fear of a possible SARS-CoV-2 infection. No other respiratory symptoms, such as a productive cough, were noted, and he denied fever. The next morning, he experienced oppressive retrosternal chest pain that lasted a few hours. Another episode of chest pain later in the day led to him presenting to the ED.Prior medical history was notable for arterial hypertension, hyperuricemia, and class D  chronic obstructive pulmonary disease with chronic hypercapnic respiratory failure. The patient quit smoking 12 years before, and his last admission was two years prior for respiratory decompensation. His current medications included chlorthalidone, tiotropium bromide, formoterol, fluticasone, and aminophylline. He was also under daily oxygen and nocturnal noninvasive ventilation. A suspected allergy to ciprofloxacin was noted.The differential diagnosis included acute coronary syndrome, Takotsubo syndrome (TTS), acute myocarditis, acute pulmonary thromboembolism, and aortic dissection.At admission, the patient’s chest pain had resolved. Apart from obvious tachycardia, cardiac auscultation was normal. An electrocardiogram recorded at admission showed sinus tachycardia with 127 beats per minute, negative T waves in leads V4-6, and a discrete ST-segment elevation in the same precordial leads . Additionally, the patient had a slightly elevated level of C-reactive protein and lymphopenia. An arterial blood sample showed mild to moderate deterioration of blood gas exchange, with respiratory acidosis, and the patient was put on noninvasive ventilation. Chest radiography showed bilateral perihilar infiltrates and a mild right pleural effusion. SARS-CoV-2 testing was positive, establishing the diagnosis of COVID-19. Chest computed tomography (CT) excluded signs of pneumonia but revealed paraseptal emphysema. Transthoracic echocardiography revealed a moderate depression of the left ventricle systolic function, with left ventricular ejection fraction (LVEF) of 35%, with hypokinesis of the apex and distal segments of all cardiac walls  and were significantly dissociated from brain natriuretic peptide values, which were markedly elevated 3649 pg/mL). Echocardiographic changes progressively resolved throughout hospitalization. A transthoracic echocardiogram about two weeks later showed the normal systolic function of the left ventricle, with LVEF of 52% and normal-sized cardiac chambers (Video 649 pg/mLNo significant alterations in segmental contractibility were seen so cardiac angiography was postponed and antithrombotic drugs were suspended. Due to difficulties in weaning the patient from the ventilator, performing cardiac magnetic resonance imaging was not possible.Corticosteroids were initiated for his lung disease, with significant improvement in blood gas exchange during hospitalization. Nevertheless, frequent episodes of desaturation occurred associated with moderate-intensity activity during hospitalization. Hydroxychloroquine and azithromycin were withheld because of concerns about cardiac toxicity. The patient was discharged after 26 days. He had two negative sets of nasopharyngeal swabs for SARS-CoV-2 prior to discharge, therefore fulfilling the criteria for cure defined by the hospital.No readmissions were observed during the following three months.We report the case of a 72-year-old man with COVID-19 who was admitted to the hospital with symptoms of TTS. TTS, which is also known as stress cardiomyopathy or broken heart syndrome, was first described in 1990 and was named based on ventriculography findings that resembled octopus-trapping pots in Japan . TTS is The discussion of physiological mechanisms of diseases potentially related to COVID-19 is complex because distinguishing between random cardiac manifestations or myocardial lesions induced by the virus is difficult. However, acute lung diseases that cause respiratory failure, such as infections, are known potential triggers for stress cardiomyopathy . SeveralThe authors present the case of an elderly man diagnosed with COVID-19 and subsequently with echocardiographic findings suggestive of TTS, directing us to a correlation between these two entities. Not only is a possible viral tropism for cardiomyocytes highlighted, but also the contribution of severe chronic lung disease, severe respiratory failure, and significant anxiety as risk factors for the development of TTS.This is a complex situation and awareness has increased regarding the cardiovascular and pulmonary sequelae of SARS-CoV-2 infection. Several studies are currently investigating laboratory and imaging markers. Their results should help determine whether acute manifestations of COVID-19 have negative impacts on cardiopulmonary function, which seems to be the likely case.


People consume and abuse alcohol for varied reasons. Problematic alcohol use is associated with mental and physical health risks, while people exposed to multiple traumas may be more vulnerable to abusing alcohol.To evaluate the prevalence and predictors of self-reported alcohol abuse among residents of Fort McMurray and explore the correlates of self-reported alcohol abuse with some mental health conditions.A cross-sectional study adopted an online questionnaire. Sociodemographic data, trauma exposure, and clinical characteristics were collected to identify the predictors of self-reported alcohol abuse. Data were analyzed using SPSS version 25 using cross-tabulations and logistic regression analysis.2 = 4.783; p = 0.033) and anxiety symptoms , and suicidal ideations or thoughts of self-harm .Two hundred and forty-nine individuals received the survey link, of which 186 completed the survey, with a response rate of 74.7%. Most participants were females exposed to COVID-19 and either wildfire or flooding traumas. The prevalence of self-reported alcohol was 27.4%. Participants who desired mental health counselling were likely to self-report alcohol abuse . There was a significant association between self-reported alcohol abuse and self-rated moderate to high depression (X Self-reported alcohol abuse is correlated with suicidal ideations, the desire to receive mental health counselling, and anxiety and depression symptoms. Therefore, initiatives to minimize mental health disorders are crucial to reducing alcohol abuse and promoting health among vulnerable populations.None Declared


Ultra-short 230 fs laser pulses of 515 nm wavelength were tightly focused into 700 nm focal spots and utilised in opening ∼400 nm nano-holes in a Cr etch mask that was tens-of-nm thick. The ablation threshold was found to be 2.3 nJ/pulse, double that of plain silicon. Nano-holes irradiated with pulse energies below this threshold produced nano-disks, while higher energies produced nano-rings. Both these structures were not removed by either Cr or Si etch solutions. Subtle sub-1 nJ pulse energy control was harnessed to pattern large surface areas with controlled nano-alloying of Si and Cr. This work demonstrates vacuum-free large area patterning of nanolayers by alloying them at distinct locations with sub-diffraction resolution. Such metal masks with nano-hole opening can be used for formation of random patterns of nano-needles with sub-100 nm separation when applied to dry etching of Si. Surfaces are fundamentally important for applications in (photo)catalysts, defrosting coatings, anti-bacterial, and biocidal nano-texturing of surfaces, as well as anti-reflection coatings. In most of these cases, the texture is key to the improved performance. However, materials and their composition are another important parameter, which has to be controlled. Surface properties can be also be chemically altered to increase toughness, e.g., Gorilla glass (Corning in 2007) for increased screen durability for mobile phones or handheld computing devices. Another example of tailoring mechanical properties was recently demonstrated in a single-crystal CoThe development of intermetallic compounds and alloys has significantly extended the range of properties of metallic materials . Nowaday2 , CuPd  and perchloric acid HClO4 etchant was utilised . SiO2 is not affected by the Cr-etch.Chromium was deposited by electron beam evaporation. For its removal, a commercial Cr-etch  based on Ceric ammonium nitrate (NHFemtosecond (fs-)laser microfabrication setup based on Pharos (Light Conversion) fs-laser was integrated with scanning Aerotech xy-stages and software control of laser emission and scanning conditions (Workshop of Photonics).Scanning electron microscopy (SEM) was used for structural characterisation of samples processed by laser and plasma treatments (Raith 150TWO electron beam writer was used in field-emission SEM mode). For the elemental analysis, the X-ray energy dispersive spectroscopy (EDS) was carried out with INCA X-act  setup on a Supra 40VP  SEM instrument.2O3 films of tens-of-nanometre thickness was recently demonstrated for photonic crystal (PhC) patterns on a large area , which is also kinetically enhanced at high temperatures. A detailed study of surface chemical composition by X-ray photo-electron spectroscopy (XPS) and Raman (for oxides) is required for identification of the compounds formed.Parameter study of mask writing by nano-hole ablation shows a different morphology of the ablation area. At pulse energies above ∼10 nJ, irregular ablation craters were formed with molten crown-type splash morphology, while at lower pulse energies, a clear circular rim was formed b. Prolonmaterial . Cr-rich3 is water soluble . It was demonstrated that even low 3 at%. concentration of Si in Cr makes oxidation and volatilization of oxides, as well as nitridation, much slower  is the surface recombination velocity.Chromium and gold  were used for comparison of light localisation and enhancement . Obvioussurfaces . HydrogeThe same boundary condition discussed above is applicable to the conductive ring and defines the local normal E-field according to the condition It is shown that alloying a nano-film with a substrate (underneath it) can be achieved with precise control of energy deposition during ablation, when nano-ring structures are formed, and a controlled remelting forms nano-disks below the ablation threshold. This adds another dimension to laser nano patterning and complements physical nano-texturing. Nano HEA alloys have previous been produced using laser-synthesis as nanoparticles . At the no-disks . It is s


Bispecific T-cell engagers (BiTEs) and bispecific antibodies (BiAbs) have emerged as novel therapeutic modalities in the treatment of advanced hematological malignancies. BiTEs and BiAbs redirect T cells to attack tumors and facilitate T-cell-mediated cell death. Blinatumomab was the first BiTE to display proof-of-concept with its remarkable contribution towards the treatment of acute lymphoblastic leukemia. Nearly a decade later, several BiTEs/BiAbs targeting a range of tumor-associated antigens have transpired in the treatment of multiple myeloma, non-Hodgkin’s lymphoma, acute myelogenous leukemia, and acute lymphoblastic leukemia. This review summarizes the most recent evidence emerging from clinical trials regarding the use of BiAbs and BiTEs in hematological malignancies whilst highlighting the limitations of these therapeutic options and providing practical insights towards overcoming these limitations.Bispecific T-cell engagers (BiTEs) and bispecific antibodies (BiAbs) have revolutionized the treatment landscape of hematological malignancies. By directing T cells towards specific tumor antigens, BiTEs and BiAbs facilitate the T-cell-mediated lysis of neoplastic cells. The success of blinatumomab, a CD19xCD3 BiTE, in acute lymphoblastic leukemia spearheaded the expansive development of BiTEs/BiAbs in the context of hematological neoplasms. Nearly a decade later, numerous BiTEs/BiAbs targeting a range of tumor-associated antigens have transpired in the treatment of multiple myeloma, non-Hodgkin’s lymphoma, acute myelogenous leukemia, and acute lymphoblastic leukemia. However, despite their generally favorable safety profiles, particular toxicities such as infections, cytokine release syndrome, myelosuppression, and neurotoxicity after BiAb/BiTE therapy raise valid concerns. Moreover, target antigen loss and the immunosuppressive microenvironment of hematological neoplasms facilitate resistance towards BiTEs/BiAbs. This review aims to highlight the most recent evidence from clinical trials evaluating the safety and efficacy of BiAbs/BiTEs. Additionally, the review will provide mechanistic insights into the limitations of BiAbs whilst outlining practical applications and strategies to overcome these limitations. T-cell-redirecting strategies have emerged as highly promising therapeutic modalities for the treatment of hematological malignancies. Notably, two approaches, namely chimeric antigen receptor T cells (CAR-T) and bispecific antibodies (BiAbs), have shown remarkable efficacy in the treatment of hematological malignancies. CAR-T cells have revolutionized the management of relapsed and refractory hematological malignancies like multiple myeloma (MM), non-Hodgkin’s lymphoma (NHL), and acute lymphoblastic leukemia (ALL) ,2,3,4. DOn the other hand, BiAbs and bispecific T-cell engagers (BiTEs) offer the T-cell-redirecting capabilities of CAR-T as an off-shelf therapy whilst eliminating the logistical and time constraints associated with CAR-T delivery. Additionally, BiAbs and BiTEs appear to have a more favorable safety profile, with lower incidences of CRS and neurotoxicity than CAR-T therapy .The pioneering success of blinatumomab as the first BiTE demonstrated proof-of-concept evidence with its remarkable contribution towards the treatment of ALL . Nearly Given that our focus is summarizing clinical data on the use of BiAbs in hematological malignancies, a detailed discussion of the basic science of these drugs is beyond the scope of this review and we direct readers towards other reviews that have explored this area in greater detail ,10.CAR-T therapy involves engineering a T cell to express a chimeric antigen receptor (CAR) specific to a tumor-associated antigen. Administering CAR-T cells would hence augment the anti-tumor immune response. BiAbs achieve a similar goal by containing two binding sites, enabling them to bind two epitopes on the same antigen or two different antigens ,11. One Like other modalities of cancer treatment, tumors can become resistant to BiAbs/BiTEs and consequently impair therapeutic efficacy. The administration of BiAbs imposes significant selection pressures on tumor clones expressing the target tumor-associated antigen, but inadvertently confer a selective advantage to sub-clones lacking the target antigen, resulting in their expansion and resistance . To counThe introduction of protease inhibitors, immunomodulatory drugs (IMIDs), and anti-CD38 monoclonal antibodies has significantly improved multiple myeloma (MM) patient outcomes , but thoA prime target for BiTE therapy in MM is the B-cell maturation antigen (BCMA). BCMA is selectively expressed on the surfaces of plasma cells and is associated with disease severity and unfavorable prognostic outcomes ,30. TeclThe use of BCMA-targeting therapeutic modalities in MM may result in either the decreased or complete loss of BCMA expression on MM cells and consequent antigen escape . To addrThe Fc Receptor Homolog 5 (FcRH5) and CD38 have been considered potential targets for BiAbs in MM . CevostaThese findings indicate that monotherapy with BiAbs holds tremendous therapeutic potential in MM patients . HoweverBesides antigen escape, the immunosuppressive TME in MM poses challenges to the efficacy of BiAb therapy. The immune microenvironment in MM is characterized by the infiltration of T-regs and the upregulation of programmed death ligand 1 (PD-L1) on MM cells ,48,49. TReprogramming the TME to augment anti-tumor T-cell immunity may also improve MM responses to BiAb. In this context, IMiDs, such as lenalidomide and thalidomide, have also demonstrated the ability to enhance T-cell-directed responses against MM cells in vitro and in vivo . DaratumIn summary, BiAbs represent an effective therapeutic approach for RRMM in terms of response rates and safety profiles. Overcoming antigen escape, either by combinatorial approaches employing mechanistically diverse BiAbs or pharmacologically upregulating the expression of target antigens, is an emerging area of investigation. Overcoming the immunosuppressive TME, either by earlier intervention or reprogramming it through IMiDs or ICIs, holds promise in terms of improving outcomes in RRMM patients.Precursor B-cell acute lymphoblastic leukemia (B-ALL) is characterized by the malignant proliferation of B-lineage precursor cells in the bone marrow and peripheral blood . While sCD19, a critical mediator of B-cell signaling, is expressed on most cells of the B-lymphocyte lineage and is maintained during the neoplastic transition of precursor B cells in B-ALL . This maThe success of blinatumomab has inspired investigations into its potential utility across different clinical presentations in B-ALL. Blinatumomab has been studied as an adjunct to consolidation chemotherapy in patients with MRD-negative B-ALL, demonstrating significant improvements in overall survival compared to standard consolidation alone . It has In summary, revolutionary advances in B-ALL treatment have been made, spearheaded by the development of blinatumomab. Lastly, the immunosuppressive TME in ALL may promote resistance to blinatumomab . A higheNon-Hodgkin’s lymphoma (NHL) encompasses a diverse group of lymphoproliferative neoplasms with varying grades of progression and severity . Among tBiAbs targeting multiple effector cell surface markers  and B-cell antigens  have been developed for NHL treatment . CD20 isRegarding adverse events, low-grade CRS is the most common side effect associated with CD20xCD3 antibodies, with grade ≥3 occurring rarely . CRS eveCD19 is another potential target antigen in NHL due to its ubiquitous expression on B cells, including neoplastic B cells . CD19-taThe CD20xCD3 BiAbs mosunetuzumab, glofitamab, and epcoritamab have received accelerated FDA approval for specific NHL subtypes due to their substantial efficacy. However, factors impairing the efficacy of CD20xCD3 BiAbs require further exploration. For example, antigen escape resulting from reduced CD20 expression has been observed in a significant number of NHL patients treated with rituximab and was associated with an inferior prognosis . Loss ofChronic exposure to BiAbs results in continuous T-cell stimulation and subsequent exhaustion, impairing the efficacy of BiAbs/BiTEs . In vitrAcute myelogenous leukemia (AML) is characterized by the infiltration of immature leukemic cells in the bone marrow and their accumulation in peripheral blood . SurvivaCurrent bispecific T-cell engagers in AML target many antigens, including CD33, CD123, CLL-1, and FLT-3  149]. C. C149]. CD123, the low-affinity binding subunit of the IL-3 receptor, has emerged as another target antigen for BiTE therapy in AML. CD123 is widely expressed on leukemic stem cells and myeloid blasts and correlates with disease severity and therapy resistance ,156,157.CLL-1 (CLEC12A) is another potential target antigen for BiTE/BiAb therapy in AML due to its expression on leukemic stem cells and myeloid blasts but not hematopoietic stem cells ,162,163.BiTEs hold promise in AML treatment, but clinical trials are currently in their infancy . There aThe previous sections have expounded on the therapeutic potential of BiAbs/BiTEs in hematological malignancies. However, ensuring the safety and tolerability of these therapeutic modalities is of paramount importance to incorporate them into treatment protocols for hematological malignancies. The primary adverse events associated with BiAb and BiTE treatment include CRS, infections, hematological toxicity, and neurotoxicity . This seCRS is characterized by an exaggerated inflammatory response with elevated levels of cytokines interleukin-2 (IL-2), interleukin-6 (IL-6), interferon-gamma (IFN-γ), and tumor necrosis factor-alpha (TNFα) ,187. TheTherefore, intensive monitoring is essential for patients receiving BiAb/BiTE therapy, particularly during the initial 48 hours of dose administration. Step-up dosing has been shown to mitigate the release of inflammatory cytokines and reduce the duration and intensity of CRS . AdditioWhen patients develop CRS, immediate supportive care is needed, followed by admission to intensive care units. Supportive care includes the maintenance of normoxia and administration of fluids/antipyretics. Furthermore, the administration of steroids and/or IL-6-blocking mAb tocilizumab can significantly alleviate both the duration and severity of CRS .It is vital to closely monitor patients receiving BiAb/BiTE therapy for signs of infection. Patients should be screened for opportunistic or reactivation infections such as cytomegalovirus and Epstein–Barr virus . A poolePatients treated with BiAbs/BiTEs are predisposed to the development of hematological toxicities, including anemia, thrombocytopenia, and neutropenia. The exact mechanism behind the development of these toxicities remains unknown but may be linked to therapy-induced pro-inflammatory cytokine release and/or the impairment of hematopoiesis ,199. SupNeurotoxicity in the context of BiAb/BiTE therapy can arise either from CRS or as a consequence of immune effector cell-associated neurotoxicity syndrome (ICANS) . The symBiAbs/BiTEs have transformed the treatment landscape for advanced hematological malignancies, with several approved BiAbs/BiTEs showing promising efficacy and favorable safety profiles. Additionally, early-stage clinical trials of numerous other BiTEs have demonstrated encouraging anti-neoplastic activity, raising optimism for their approval in the coming years.However, the efficacy and tolerability of certain BiAbs/BiTEs warrant further exploration. Antigen escape is a major resistance mechanism to BiAb/BiTE therapy. Elucidating additional tumor-associated target antigens and exploring combinatorial, multi-antigenic BiAb/BiTE may counteract antigen escape. Additionally, the immunosuppressive TME in hematological malignancies is a significant contributor to BiAb/BiTE resistance. Investigating strategies to normalize the tumor microenvironment, such as immune checkpoint inhibitors and immunomodulatory agents, may enhance the efficacy of BiAbs/BiTEs and mitigate therapeutic resistance.Presently, much of the BiAb/BiTE research in hematological malignancies focuses on patients with relapsed/refractory disease, but it is plausible that patients in the early stages of their disease may respond to BiAbs/BiTEs more favorably due to their lower tumor burden and a TME more conducive to anti-tumor immune responses. We have already discussed how T-cell dysfunction consequent to exposure to multiple lines of cancer therapy has been associated with resistance to BiABs, early-stage disease being associated with a better therapeutic response to BiTEs, and clinical trials being underway to assess the efficacy of BiAbs/BiTEs in early-stage hematologic malignancies. Exploring novel BiAb engineering strategies such as trispecific antibodies—which target more tumor antigens and minimize the risk of antigen escape—is also important. Incorporating natural killer cell engagers may also synergize with bispecific T-cell engagers and potentially enhance the anti-tumor immune response.In conclusion, the field of bispecific T-cell engagers holds tremendous therapeutic potential, and we eagerly anticipate further progress from future preclinical studies and clinical trials. The continued advancement of these therapies is expected to have a significant impact on the treatment of hematological malignancies, bringing hope to patients and healthcare professionals alike.


Nmnat pre-mRNA to regulate the outcomes of alternative splicing. However, the fundamental question remains whether such microRNA-mediated interference with RNA secondary structures is a global molecular mechanism for regulating mRNA splicing. We designed and refined a bioinformatic pipeline to predict candidate microRNAs that potentially interfere with pre-mRNA stem-loop structures, and experimentally verified splicing predictions of three different long-range pre-mRNAs in the Drosophila model system. Specifically, we observed that microRNAs can either disrupt or stabilize stem-loop structures to influence splicing outcomes. Our study suggests that MicroRNA-Mediated Obstruction of Stem-loop Alternative Splicing (MIMOSAS) is a novel regulatory mechanism for the transcriptome-wide regulation of alternative splicing, increases the repertoire of microRNA function and further indicates cellular complexity of post-transcriptional regulation.While RNA secondary structures are critical to regulate alternative splicing of long-range pre-mRNA, the factors that modulate RNA structure and interfere with the recognition of the splice sites are largely unknown. Previously, we identified a small, non-coding microRNA that sufficiently affects stable stem structure formation of  MicroRNA-Mediated Obstruction of Stem-loop Alternative Splicing (MIMOSAS) is a novel regulatory mechanism for the transcriptome-wide regulation of alternative splicing. Alternative splicing plays an important role in cellular differentiation and stress response in both physiological and pathological conditions. While the vast majority of human genes are alternatively spliced –3, 22% oSplicing of long introns is likely facilitated by RNA secondary structure formation in the intron of the pre-mRNA, bringing 5’ and 3’ splice sites into relatively close spatial proximity and promoting excision of a particular intron. While many such long-range interactions are evolutionary conserved and are strongly associated with alternative splicing –12, a keNMNAT pre-mRNA  that have been well studied in mRNA regulation , 14, typnt that hpre-mRNA . In partpre-mRNA . Based o from RA .in vivo in Drosophila and in cultured human cell models.While our initial consideration was limited to a pair of one microRNA and mRNA, here we show that MIMOSAS is a potentially pan-genomic splicing mechanism. In particular, we designed a computational pipeline that (i) screens for potential sites that form splice-relevant stem-loop RNA structures on different genes, and (ii) computationally predicts microRNAs that disrupt such secondary structures. Most importantly, we experimentally corroborated our computational predictions Drosophila genome (DGopen = G0 − G1). We consider a microRNA candidate if the energy of the duplex binding structure between a microRNA and mRNA (Gduplex) is larger than DGopen, indicating that the binding of the microRNA that destroys the splice-relevant secondary structure is energetically more favorable (Gduplex) and opened structure (DGopen), DGMIMOSAS = Gduplex − DGopen, as well as the probability that a microRNA binds a given box  when we determined the energetically most stable structure of the underlying mRNA subsequence. Utilizing these box sequences in DGMIMOSAS and chose two microRNAs that target the underlying boxes energetically (un)favorably. We expected that identified candidate microRNAs interfere with the formation of the stem structure, driving the RC splice variant, while we expected splice variants RA/B/D/E/F/G in the absence of MIMOSAS  splice variant, while we observed no significant changes with miR-976 or miR-1000 over-expression  and confirmed that the corresponding stem almost completely overlapped with the boxes we found through complementary sequences. In particular, when the boxes form a stem, the RA splice variant is produced where exon 5 was spliced out. In turn, the RB splice variant emerges when the stem does not form, leading to a retained exon 5 . As for miR-137, we observed a strong binding probability with the sequence of box I, while we found no binding signal when we considered miR-307a. Furthermore, we considered miR-278 and miR-983 that showed strong probabilities binding box II . To corroborate our predictions, we performed real-time PCR after expressing corresponding microRNAs or microRNA sponges to reduce endogenous microRNAs. Notably, we found that microRNAs with favorable binding characteristics such as miR-210, miR-137 (targeting box I) as well as miR-983, miR-9b, miR-278 and miR-1002 (box II) drove the emergence of the MIMOSAS-specific RB splice variant. In turn, we found that miR-307a and miR-193 that displayed minimal binding qualities showed no effect in RB production .Previously, we found that miR-1002 potentially binds and interferes with a splice-relevant stem loop formation on the A and RB . Notablyd exon 5 . NotablymiR-1002 . Specifiin vivo. Building upon our previously published alternative splicing reporter for Drosophila Nmnat  and mCherry (STEM) signals in the brains with the co-expression of miR-210, miR-9b, miR-983, and miR-278 compared to the non-targeting (miR-9c) or negative (miR-307a) control . However, when we determined the most stable structure capturing exons 3 and 4, we observed that box II forms a different stem with a newly identified box III in exon 4 . As a consequence of the formation of splice-relevant stems between boxes I and II, or boxes II and III, Rpl3 pre-mRNA can generate additional splice variants. Specifically, formation of a stem between boxes II/III leads to RA/H production, formation of a stem between boxes I/II leads to RG, and an open confirmation without any stems leads to RD .Both n exon 3 , 11. By ds to RD . NotablyRpL3 and interfere with the stem formation of its pre-mRNA . To quantitatively measure the ratio of different RpL3 mRNA variants after microRNA overexpression, we used a two-color multiplex real-time PCR assay with specific TaqMan probes. In particular, we designed probes for the total mRNA variants, RA/H and RG, and RG only  two color reporter construct that mimics the intricate secondary structures of the pre-mRNA. We retained all the crucial sequences involved in the formation of stems in the pre-mRNA and employed self-complementing split FP  and cytoplasmic (NES) localization signals, we distinguished each spliced mRNA variant based on its color and cellular location . While microRNAs are mostly studied in the context of mature mRNA 3’UTR binding and target transcript downregulation, MIMOSAS predicts the binding of microRNAs to interior intronic or exonic regions in the pre-mRNA of targets and regulating mRNA splicing. The outcome of such microRNA-pre-mRNA duplex binding allows mRNA expression in an efficient and splice variant-specific manner. Given the dynamic and cell type/tissue-specific expression of microRNAs, MIMOSAS presents a potential new regulatory layer through which cells in different tissues can specifically control the expression of transcript variants. MIMOSAS also unveils regulation of gene expression by noncoding RNAs in a stress or disease state-specific manner and underscores the importance in considering alternative splicing modulation as a therapeutic means. As 60% of point mutations associated with diseases have been found to be related to splicing defects (Here, we predict candidate microRNAs that directly interfere and drive alternative splicing through binding with pre-mRNA stem-loop structures. Providing testable hypotheses through a computational pipeline, we experimentally verified the splicing predictions of three different long-range pre-mRNAs in mammalian cell culture models and in vivo in  defects , several defects , and a m defects . Given t


Problematic substance use is rising, and other mental health conditions like anxiety and depression correlate with substance abuse. Diverse interventions to reduce this effect are emerging. Supportive text messages offer the prospect of improving symptoms of drug misuse and other associated comorbidities.The study aims to evaluate the impact of the Text4Hope-Addiction program in mitigating craving, anxiety, and depression symptoms in subscribers.Individuals self-subscribe to Text4Hope Addiction program by texting “Open2Change” to 393939 to receive daily addiction-related text messages for three months. Subscribers are invited via text message to complete online questionnaires which assess cravings, anxiety, and depressive symptoms using the Brief Substance Craving Scale, Generalized Anxiety Disorder-7 Scale, and Patient Health Questionnaire-9 on subscription (baseline), six weeks and three months. Data were analyzed using SPSS version 25 with descriptive and inferential statistics. Satisfaction responses were used to assess various aspects of the Text4Hope- Addiction program.There was a significant difference in the mean baseline and three-month BSCS scores , PHQ-9 scores , and the GAD-7 scores . Participants agreed that the supportive text messages helped them cope with addiction-related stress (89%), anxiety (81%) and depression (69%).The Text4Hope-Addiction program effectively reduced cravings, anxiety, and depression among subscribers, with high satisfaction rates for the program. Healthcare practitioners and policymakers should consider implementing supportive text-based strategies to complement conventional treatments for addiction.None Declared


EGFR and KRAS oncogenes, which are detrimental to cancer cells when combined. The co-expression of mutant KRAS and EGFR potentiates MAPK signaling through extracellular-signal-regulated kinases (ERK1/2), which mediate toxicity, thereby inducing morphological changes and increased micropinocytosis in lung adenocarcinoma cells [Cancers features five new articles, four of which focus on EGFR-mutant NSCLC patients, while one reports on the characteristics and clinical outcomes of Norwegian KRAS-mutant NSCLC patients.Genetic mutations can activate different sets of proto-oncogenes and tumor suppressors genes. However, in lung adenocarcinoma, some combinations of mutations are mutually exclusive, such as mutations in nases ERK/2, whichEGFR-mutant NSCLC cells. The researchers [EGFR sensitized with T790M/C797S amplify ERK signaling and that these cells are sensitive to trametinib, an MEK inhibitor. Christine M. Lovly’s group [EGFR-mutant cell lines. Furthermore, the addition of BKM 120 (a PI3K inhibitor) or dasatinib (an SFK inhibitor) enhanced the effects of Osimertinib. Likewise, a dasatinib–Osimertinib combination was the most effective at inhibiting tumor cell proliferation. Src family kinases (FAK) have proven to be relevant in Osimertinib-resistant EGFR-mutant tumor cells [EGFR-mutant cells produce high IL-6 levels and that the blockade of the IL-6/gp130/JAK pathway led to a decrease in phosphorylated STAT3 levels, but not when dasatinib was used, indicating that the combination of STAT3 and Src/SFK inhibition could be necessary [EGFR-mutant NSCLC cells that are becoming resistant undergo an epithelial-to-mesenchymal transition (EMT), wherein IL-6 continues to be expressed independently of the activation of EGFR and under the control of TGF-β [EGFR-mutant cells. In the related study, treatment with IL6 antibodies enhanced antitumor immunity and the efficacy of immune checkpoint anti-PD1 inhibitor [EGFR mutations suppress TNF mRNA levels by inducing the expression of miR-21, which leads to a decrease in TNF mRNA levels. Conversely, EGFR inhibition  leads to a loss of miR-21 and an increase in TNF mRNA levels and, consequently, TNF-induced NF-kB activation [We-Chien Huang and colleagues  identifiearchers  observed’s group  previousor cells ,5,6, whior cells , whereinnhibitor . Controvtivation .EGFR inhibitors. They found that the loss of NFKB1A, which suppresses NF-kB signaling, is associated with poor progression-free survival. Furthermore, suppressing the NF-kB pathway enhances the response to EGFR TKIs in EGFR-mutant models [EGFR and TNF (etanercept) could increase the therapeutic benefit in EGFR-mutant NSCLCs. Therefore, the aforementioned study by Huang et al. [EGFR-mutant tumors and highlights the need to further understand ERK signaling and the role of miR-21 in light of the apparently divergent findings of Huang et al. [Trever G Bivona and colleagues reported that NF-kB signaling causes resistance to t models ,13. Gongt models  suggesteg et al. . EGFR-mutant NSCLCs. The identification of these factors raises tantalizing questions. However, prior to the discovery of EGFR mutations, Rafaella Sordella and colleagues had already identified that adequate treatment for mutant-EGFR requires pharmacological inhibitors of the signal transducer and activator of transcription (STAT3) and Akt signaling pathways [According to the study conducted by Huang et al. , IL-6 plpathways .Cancers, Yan-Jei Tang and colleagues [EGFR T790M mutation instead of administering Osimertinib as a first-line treatment for EGFR-mutant NSCLCs. This is a complex dilemma that involves subtle intricacies in clinical management and underlying biological mechanisms. According to the ASCO Living Guidelines, EGFR-mutant patients  with PS0-2 should be treated with Osimertinib [EGFR-mutant NSCLCs with exon 19 deletion presented better survival than those with the L858R mutation [EGFR-mutant patients with an exon 19 deletion and brain metastases present better survival with Osimertinib compared to other EGFR TKIs [Also included in this Special Issue of lleagues  propose mertinib . Howevermutation . AdditioGFR TKIs . The retGFR TKIs  also fouEGFR mutations, Osimertinib resulted in longer overall survival compared to gefitinib or erlotinib. However, a subgroup analysis found that this advantage was not seen in Asian patients or in patients with the L858R mutation  [The FLAURA trial clearly revealed that in patients with previously untreated NSCLC with 71–1.40) . EGFR exon 19 deletions has been associated with an enrichment in the occurrence of T790M mutations [While the lack of an overall survival advantage in Asian patients deserves further investigation to uncover its underlying genetic reasons, the advantage of Osimertinib with respect to EGFR exon L858R mutant NSCLCs is lower. Additionally, this type of mutation is often associated with TP53 mutations and 3q23 amplification, of which the latter contains the MRAS gene that is part of the SHOC2 phosphatase complex involved in resistance to EGFR TKIs [In contrast, the probability of developing T790M mutations in GFR TKIs . LikewisGFR TKIs  reinforc2+-dependent phosphatidic acid phosphatase (PAP) enzyme that converts phosphatidic acid to diacylglycerol, a precursor of triacylglycerol and phospholipids. In this study, the authors observed that gefitinib treatment induced LPIN1 expression, thereby enhancing diacylglycerol concentrations in EGFR-TKI-resistant H1650 cells followed by activating protein kinase C-δ and NF-kB. This finding sheds new light on the role of NF-kB activity in resistance to EGFR TKIs. The authors also found that the loss of LPN1 expression sensitizes EGFR-mutant NSCLC cells to gefitinib in vivo. The investigators used propranolol as an LPIN1 inhibitor [EGFR-mutant NSCLC cells. Propranolol was used as a β-blocker, which efficiently inhibited resistance to EGFR TKIs [EGFR-mutant NSCLCs [The third study in this Special Issue was conducted by Jung Hee Cho and colleagues . The invnhibitor  and attaGFR TKIs . The relGFR TKIs . Based ot NSCLCs .Cancers, Cansouline et al. report a revisitation of the position of EGFR TKIs in early surgically resected EGFR-mutant NSCLCs [EGFR-mutant stage IB-IIA NSCLC after complete resection, showing a 4-year disease-free survival rate of 73% with Osimertinib and 38% with a placebo (hazard ratio 0.27). Furthermore, it supports the use of adjuvant Osimertinib in resected EGFR-mutant NSCLCs due to improved central nervous system disease survival [In this Special Issue of t NSCLCs . The stusurvival . EGFR-mutant cells, the mechanisms of resistance mentioned earlier could also occur in the adjuvant and neoadjuvant settings. Additionally, the promigratory signal Src-homology-2 domain-containing phosphatase-2  activates Src family kinases (SFK). We previously reported that high levels of SHP2 mRNA correlated with poor progression-free survival and overall survival in metastatic EGFR-mutant NSCLC cells treated with EGFR TKIs [EGFR-mutation-positive lung adenocarcinomas but not in the EGFR wild-type [However, as seen in stage IV NSCLC patients and GFR TKIs . We alsoild-type . Ito et ild-type  reportedild-type .KRAS-mutant NSCLCs, including the different genotypes of druggable KRAS G12C and non-G12C mutations, while scrutinizing potential variables that could shed light on the effects of anticancer therapies. Wahl and colleagues [KRAS mutations is lower in these populations than in Caucasian NSCLC patients [KRAS G12C inhibitors presents new opportunities and challenges for researchers investigating the mechanisms of resistance to sotorasib, adagrasib, and SHP2 inhibitors, which appear to be analogous to those identified in other targeted therapies such as EGFR TKIs in EGFR-mutant NSCLCs [The fifth article included in this Special Issue by Sissel Gyrid Freim Wahl  shares slleagues  add furtpatients ,31. The t NSCLCs .


This systematic review and meta-analysis aims to compare the effects of couple-based prevention interventions against individual-level interventions on HIV prevention in randomized controlled trials (RCTs), identify potential moderators, and assess study quality. Eleven RCTs were included, comprising 3933 couples in the intervention group and 7125 individuals in the individual control group, predominantly in heterosexual couples from the USA and Africa. Couple-based interventions had a more significant effect in promoting condom use and HIV testing. Education levels of high school or above, residence in low- and middle-income countries, and intervention design incorporating HIV counseling and testing were associated with higher odds of condom use. The quality assessment analysis identified methodological and theoretical heterogeneity factors. Evidence of couple-based HIV prevention RCTs among men who have sex with men, injecting drug users, sex workers, and transgender women warrant further investigation. Recommendations are made to improve the quality and replicability of future intervention studies.The online version contains supplementary material available at 10.1007/s10461-022-03768-5. Sexual transmission of HIV between two sexually intimate partners remains the primary cause of the HIV epidemic in many parts of the world . In thisThe past two decades have witnessed growing advancement in couple-based HIV prevention programs. Recent empirical evidence supports the credibility of couple-based interventions, particularly when compared to individual-level interventions with large base sizes. A recent meta-analysis suggested that couple-based interventions are more effective than individual-level interventions in promoting HIV-protective behaviors to prevent HIV transmission and infection . Dyadic Parallel to this, the development of HIV-prevention randomized controlled trials (RCTs) in key populations  and their sexual partners has become an overarching public health issue. One important observation is that in recent years, more than 60% of new adult HIV cases worldwide occurred in key populations . HoweverTo date, five systematic reviews and meta-analyses of HIV couple-based intervention and prevention studies have been published, which primarily provide qualitative and quantitative assessments of key intervention elements and study outcomes , 16–18. First, the scarcity of couple-based HIV prevention interventions has led to notable heterogeneity in study designs and outcomes in these five systematic reviews and meta-analyses. Although one of the five systematic reviews and meta-analyses compares couple-based and individual-level interventions , it achiFurthermore, methodological heterogeneity in HIV prevention intervention research may also be perceived on a theoretical level. Couple-based research frequently relies on individual-level theories as design and interpretation frameworks , potentiSecond, there is distinct heterogeneity in the quality assessments used to substantiate each study’s internal validity among these five meta-studies. Only Jiwatram-Negrón and El-Bassel’s study  used an Considering the heterogeneity within the design, outcome, and quality assessment variables of previous meta-studies, this systematic review and meta-analysis aims to (a) estimate the relative magnitude of couple-based interventions over individual-level interventions’ effects on HIV preventive behaviors of sexual-risk reduction through direct comparison of RCTs; (b) identify potential moderators, particularly intervention- and relationship-specific factors of the intervention effect; and (c) assess the methodological and reporting quality of the intervention. This paper expands on existing systematic reviews and meta-analyses, integrating more recent literature with more robust methodological support and making suggestions for quality assessment.https://www.crd.york.ac.uk/PROSPERO/). We conducted and reported our systematic review and meta-analysis according to Preferred Reporting Items for Systematic Reviews and Meta-Analysis  independently searched five electronic databases . The keywords combined participant (Couple OR dyad OR partner OR married), intervention-related terms (training OR intervention OR prevention), and disease-related terms (HIV OR AIDS) (See detailed search strings in Table B in Online Attachment). We also searched the reference lists of the published systematic review and original articles. All searches were limited to English peer-reviewed journal articles.Studies were eligible if they met the following criteria: (1) used RCT study design; (2) evaluated a couple-based preventive intervention compared to an individual-level control, including biobehavioral components  as previously categorized by Jiwatram-Negrón and El-Bassel ; (3) conTwo authors (RF & JH) independently screened the results based on titles, abstracts, and full texts. RF & JH held three discussion meetings with the corresponding author (NXY) to resolve any discrepancies during the screening phase. The full texts were excluded if they 1) lacked RCT design, (2) lacked a couple-based or couple-focused intervention or control group, (3) lacked couple vs. individual comparison, (4) used the same dataset published in a previous paper, (5) used a secondary analysis, (6) provided only baseline data, (7) randomized only HIV-negative partners, (8) described study outcomes that were not related to sexual-risk reduction or HIV/AIDS, or (9) invited study participants who were not in a sexual relationship (Table C in Online Attachment). The selection process is shown in Fig.  lacked RThe outcomes of interest are HIV-preventive behaviors, including condom use (defined as consistent condom use or no condomless sex), HIV testing, STI reduction, and ART adherence. ART adherence can be measured by either a self-report scale or blood-based tests.We (RF & JH) coded the adjusted odds ratios (aOR) with 95% confidential interval, means and standard deviations (SD) in each arm before and after interventions, and mean changes with variance in each arm or other data format used to calculate odds ratio.https://shimo.im/desktop) to extract participant-related information , intervention-related data , and implementation-related characteristic .We (RF & JH) also used a Shimo spreadsheet  to estimate the comparative effect of the couple-based intervention on HIV preventive behavior. An OR higher than 1 indicated greater odds of condom use in the couple-based intervention than in the individual-level control. Forest plots illustrate the effect sizes for individual and pooled studies.Q tests and I2 to assess the significance and proportion of the between-study heterogeneity . No significant asymmetry was detected from the funnel plot .Eleven comparisons reported condom use during anal or vaginal sex acts. The composite effect size for the change of condom use between the two groups was 1.431  = 1.538, p = 0.464, I2 = 0]. No significant asymmetry was detected from the funnel plot .Three comparisons reported HIV testing rates between the two groups. The composite effect size for the change in HIV testing between the two groups was 1.308  = 3.318, p = 0.069, I2 = 69.86]. Only one study reported ART adherence between two groups with a non-significant effect size  = 4.401, p = 0.036], compared to those had received less than a high school education. Compared to skills-building, interventions incorporating an HIV counseling and testing component were more effective in improving condom use . Improvements in condom use were higher in low- and middle-income countries (LMICs) compared to studies conducted in high-income countries .Participants with an education level of high school or above showed a higher odds of condom use [Other moderators—such as age, type of couple, HIV status, study quality, intervention sessions, tailoring, and fidelity—were not significantly associated with the intervention effect of condom use Table . The numOnline Table D summarizes the methodological quality of the included studies. All the included studies provided clear information on their eligibility criteria, randomization, similar baseline, and utilized appropriate statistical analysis for between-group comparisons –38. HoweTable Different from Jiwatram-Negrón and El-Bassel’s categorization of couple-based HIV prevention and intervention biobehavioral studies , we idenThe intervention ranged from one to 14 sessions, apart from three interventions that offered a one-off CHCT session , 29, 38.This meta-analysis is the first to synthesize existing RCTs, and it finds that couple-based interventions are more efficacious in promoting condom use and HIV testing  than individual-level interventions. As moderators for intervention effect, participants with higher education levels, couple-based interventions utilizing HIV testing and counseling strategies, and interventions conducted in low-income countries showed more significant improvement in condom use. This suggests that, despite the potential to reduce sexual risks and enhance HIV testing, many couple-based HIV prevention interventions conducted worldwide remain inadequate. Therefore, the following discussion also highlights significant methodological constraints of RCTs that are critical for facilitating advancements in couple-based HIV prevention interventions.This systematic review suggests that couple-based interventions have a stronger effect than individual-level interventions to boost condom use and encourage HIV testing among couples in RCTs. One possibility is that behavioral cooperation may help couples establish common goals in initiating healthy behaviors and developing a partnership based on joint decision-making and collective action toward reducing HIV transmission risks . When boAnother possible consideration is that enhanced communicative skills may enable couples to discuss complex or private sexual issues more openly. However, such inter-couple discussions are hard to implement without proper guidance , 40. CouCommunication-based intervention techniques may help couples to acknowledge sexual consent and shift their normative perceptions of their sexual relationships , therebyWe identified three moderators of the intervention effect in promoting condom use: (1) intervention design incorporating HIV counseling and testing, (2) low- and middle-income countries, and (3) high school or above education level. First, HIV counseling and testing showed a stronger intervention effect than skills-building in promoting condom use. Although we did not conduct subgroup analyses on HIV testing due to limited comparisons, one 2020 RCT study found that the skills-building component of a couple-based intervention resulted in comparable effects on both condom use and HIV testing . It is wSecond, adapting intervention material to participants’ education levels may increase the effectiveness of HIV transmission prevention interventions. As previously mentioned, participants with a high school education or above were more likely to use condoms than those with a lower education level. Intervention research that incorporates a skills-building component and typically takes several hours of training , 32, 47 Third, couple-based HIV prevention interventions may be more beneficial to reducing sexual risks when implemented in areas where resources are more constrained, such as in LMICs. Evidence from prior research supports our findings that CHCT has been recognized as one of the most cost-effective intervention strategies in resource-constrained settings and is particularly strong in HIV-serodiscordant couples . This maSeveral methodological and theoretical barriers must be highlighted because they risk impacting the advancement of couple-based HIV prevention interventions. First, although study quality did not appear to moderate the intervention effect in RCTs, seven of the 11 studies (63.6%) failed to conceal allocation or blind the assessor. Consequently, a low-quality or inadequately designed study may still distort the verifiability of the study’s effects because of these two issues. We suggest that the allocation of quality assessment measures be executed by off-site third parties or by using sequentially numbered, sealed, opaque envelopes to establish better methodological and reporting quality.Moreover, because eight of the 11 studies (72.7%) were published before 2014, only one study reported its intervention following the TIDieR checklist for better reporting and replication . By utilRegrettably, most (six out of seven) of the reviewed couple-based HIV preventive intervention RCTs reported using theoretical frameworks were guided by individual-based theories focusing on behavioral changes at the individual level, such as social cognitive theory and combined ecological theory, rather than dyadic-level theories , 34. As We first recommend that interventions adapt a more socioculturally sensitive approach by considering both partners’ cumulative education levels in coordination with other demographic characteristics of the local target populations, such as income level and sexual orientation. Although heterosexual couples are disproportionately represented in these 11 interventions, the target participants were often subgroups of populations impacted by low income, poor housing (including homelessness), or domestic violence , 33. MorWe also suggest that interventions may benefit from more thoroughly considering relationship dynamics in a broader theoretical context. One way to do this is to identify appropriate outcome measures based on the couple’s interdependence within dyadic coping perspectives . To dateAnother method is to explore theories that integrate relationship dynamics and HIV prevention. There is a large gap between theories construing the relationship dynamics that may influence the transformation of motivation and health behavior, especially among same-sex couples in an HIV-serodiscordant relationship. A recent qualitative study revealed how “viral load agreements” facilitate the practice of different strategies in undetectable viral load for prevention among gay male couples , sheddinThere are some limitations to this study. First, a relatively small number of RCTs are available, limiting the statistical power for subgroup meta-analysis and meta-regression. Second, because all 11 studies were conducted in the USA and Africa, the conclusions drawn in this study may not be generalizable to, or adequately representative of, other world regions. Third, given that most studies have been exclusively targeted at heterosexual couples, there is an urgent need for more robust bodies of evidence on the effect of intervention studies on more diverse populations. In the future, with more original RCTs assessing intervention effects among key populations from more global perspectives, an updated meta-analysis would provide more evidence of the sustainable impact of couple-based interventions.In this systematic review and meta-analysis, couple-based interventions are more efficacious than individual-level interventions in biobehavioral HIV prevention. The intervention effect of couple-based HIV prevention RCTs will be improved by considering sociocultural sensitivities and theoretical contexts in relationship dynamics. Finally, couple-based HIV prevention RCTs are still in their infancy, and studies among key populations  warrant further investigation.Supplementary file1 (DOCX 52 KB)Below is the link to the electronic supplementary material.