The field of targeted protein degradation (TPD) is a rapidly developing therapeutic modality with the promise to tame disease-relevant proteins in ways that are difficult or impossible to tackle with other strategies. While we move into the third decade of TPD, multiple degrader drugs have entered the stage of the clinic and many more are expected to follow. In this review, we provide an update on the most recent advances in the field of targeted degradation with insights into possible clinical implications for cancer prevention and treatment. Cancer is a major public health problem. Each year nearly 20 million new cases are diagnosed and almost 10 million deaths from cancer are registered . To dateInspired by the fact that cells employ the ubiquitin-proteasome system (UPS) to maintain intracellular proteostasis, interest has focused on an alternative approach that aims to control protein function by regulating expression levels of a target protein rather than its activity. Such efforts have evolved from the discovery that proteasome inhibitors that block protein degradation have tumoricidal activity. Two examples approved for the treatment of multiple myeloma are carfilzomib and bortezomib . Newer sIn this review we will reflect on different targeted protein degradation approaches and focus on the current status of clinical translation of TPD in cancer treatment.Targeted protein degradation (TPD) has received a lot of attention as a novel and innovative chemical tool and therapeutic modality. By harnessing protein degradation pathways, TPD mediates depletion of a protein of interest from within or outside the cell , Table 2The ubiquitin-proteasome system (UPS) is an essential pathway in the cell that processes the ablation of misfolded or damaged proteins involved in the pathogenesis of different diseases . This tiActive ubiquitination and degradation of tumor suppressors  are at the root of maintaining cancer cell proliferation. Here, targeted protein stabilization (TPS) instead of degradation would prove therapeutically beneficial . RecentlThe PROTAC technology has experienced three generations of development. The first generation required peptide sequences for E3 ligase recognition and cell penetration. The first PROTAC was applied in Xenopus egg extract to target methionine aminopeptidase 2 to the Skp1-Cullin-F box complex for ubiquitination and proteasomal degradation . The samPROTACs have potential advantages compared to other traditional approaches to suppressing a POI, providing them with a greater chance for faster clinical development against a wider range of targets in multiple cancer types  37,38].,38.37,38Unlike small-molecule inhibitors that function via binding to catalytic or allosteric sites, PROTACs derived from a binder  on any POI surface pocket with reasonable affinity may cause target destruction ,38. ThisPROTACs are also able to eradicate mutated targets . In facttrans-cyclooctene-tagged POI ligand as the precursors [trans-cyclooctene, forming a covalent six-membered ring moiety, these two precursors created integrated PROTACs (CLIPTAC) in cells that successfully depleted oncogenic BRD4 or ERK1/2 in a CRBN E3 ligase- and proteasome-dependent manner [Although protein degradation has revolutionized drug discovery platforms, current PROTACs come with several shortcomings that are likely to restrict their potential as therapeutics. These heterobifunctional molecules require linker optimization, possess high molecular weight and a high polar surface area that is normally associated with poor cellular penetration, solubility and other drug-like properties. An alternative has been proposed using the idea of CLIPTACs (in-cell-click-formed proteolysis targeting chimeras) in which two small precursor molecules with the ability to click intracellularly will pass through cellular membranes more easily than one large compound . Heightmecursors . Via thet manner . The maiHigh bioavailability can be achieved by orally bioavailable prodrug PROTACs derived from CRBN ligands . Wei et Since PROTACs operate in a catalytic manner and enable systemic protein knockdown, their off-target toxicity is a major concern. One strategy to circumvent systemic undesired effects involves the use of designed peptides, so-called phosphoPROTACs, that can be conditionally activated via phosphorylation by specific growth-factor stimuli . A recenOther approaches for tissue/cell specific degradation of POIs by PROTAC while avoiding deleterious effects elsewhere employ light stimuli ,60. OptiConventional small-molecule PROTACs generally display unfavorable pharmacokinetics and lack of tumor specificity, which may contribute to systemic toxicity due to their nonspecific distribution in normal tissues. One way to achieve tumor specific delivery of PROTACs has been proposed by using gold nanoparticle (GNP)-based multi-headed PROTACs . Cer/PomSelective delivery of a broad spectrum PROTAC into specific cell types is also feasible by an antibody-PROTAC conjugate. Maneiro et al. described a trastuzumab-PROTAC conjugate in which E3 ligase-catalyzed degrader activity is caged with an antibody linker which can be hydrolyzed upon antibody-PROTAC internalization, releasing the active PROTAC and stimulating protein destruction  Figure . Proof oThe selection of appropriate E3 ligase/E3 ligand systems is crucial for the progress of PROTAC research. Application of PROTACs in an endogenous setting is mainly limited to target proteins with available ligands. An advancing solution has been provided by tag-directed chemical degrader systems in which the tag-POI fusion protein was expressed in cells and the universal PROTAC was administered to attract the candidate E3 ligase and the tag of the tag-POI fusion . MeasuriBD1L94V along with the corresponding CRBN-based heterofunctional degrader utilizing a bump-and-hole approach [BD1L94V system and the dTAG system. This strategy complements currently available degradation tags to attack disease co-dependencies.The dTAG system uses PROTACs to degrade target proteins that have been genetically fused to a mutant isoform of FKBP12 ,76. Bothapproach . They alFurthermore, Veits et al. recently reported a novel degradation methodology on fusing a POI to the small protein MTH1 (MutT homolog-1), which serves as a ligand-binding tag .This AchUnfolded or misfolded proteins expose hydrophobic regions that signal for recruitment of E3 ligases to degrade unwanted POIs in a proteasome-dependent fashion. Several studies have mimicked protein unfolding by labeling specific targets with low molecular weight hydrophobic tags to promote their destruction ,83,84,85The human genome comprises approximately 1000 predicted E3 ubiquitin ligases that are important not only in normal cellular physiology but also in diseased states, making them attractive targets for drug discovery. Overexpression of E3 ligases is associated with poor clinical prognosis or drug resistance in cancer cells. These enzymes do not exhibit deep and druggable active sites for binding to small molecules; their inhibition is generally achieved by targeting protein-protein interactions. Only few potent compounds have been described that bind to the E3 substrate recognition site. However, competition with high-affinity endogenous substrates may increase unspecific cytotoxicity. Further, E3 ligases are multi-domain and multi-subunit enzymes . Thus, tRecently, He et al. extended this principle to MDM2, an oncogenic E3 ligase and antitumor target . A homo-RNA-PROTACsA new type of PROTAC, so-called RNA-PROTAC, aims to degrade RNA binding proteins (RBPs), a class that until now has been proven difficult to target pharmacologically . RBPs inTranscription Factor PROTACsTranscription factors (TF) are central in numerous diseases, yet remain incurable due to the lack of enzymatic activity and ligandable sites. In a recent study, Crews and colleagues have co-opted the DNA-binding capability of TF to develop TRAnscription Factor TArgeting Chimeras (TRAFTACs) . The TRAA complementary drug discovery and development platform has been proposed by the O-PROTAC model, in which a double-stranded oligonucleotide is incorporated as a TOI binding moiety in PROTAC  Figure b. This mThe field evolves at a rapid pace as highlighted by a recent report of TF-PROTAC that links an DNA oligonucleotide to an E3 ligase ligand via a click reaction, to selectively degrade the TOI c. Here, Altogether, compared with RNA, the DNA oligomer is more stable and the DNA binding specificity of TFs is better defined than the RNA binding specificity of RBPs. Therefore, TF-targeting platforms expand the druggable target spectrum with therapeutic benefits for patients with cancer and other diseases.G4-PROTACsG-quadruplexes (G4s) are four-stranded nucleic acid structures of DNA or RNA rich in guanine bases that are enriched in gene promoter regions. Many G4s harbor physicochemical and structural properties that render them favorable for drug design. Transcriptional repression of pathogenic proteins through stabilization of G4 structures or telomerase inhibition by telomeric-G4s have been suggested as novel antitumor strategies; a first-in-class G4-interacting compound has reached phase II trials in neuroendrocine/carcinoid tumors (NCT00780663) . RecentlAptamer-PROTACsAptamers are single-stranded DNA or RNA oligonucleotides that bind to the target protein with high specificity and affinity. They possess favorable in vivo safety profiles without potential immunogenicity, establishing them as targeted therapeutics in oncology ,107,108.Recently, Tan and colleagues provided the first proof of concept evidence using nucleic acid aptamer as a targeting ligand . The desMolecular glue degraders are small, drug-like monovalent compounds that induce interactions between an E3 ligase and a target protein, promoting the degradation of the latter in a proteasome-dependent manner  Figure . Unlike Similarly, the clinically approved thalidomide and its analogs known as immunomodulatory drugs (IMiDs) exert their therapeutic activity via a molecular glue mechanism by reprogramming the target spectrum of the E3 substrate receptor CRBN. IMiD binding to CRBN resulted in proteasomal depletion of the IKAROS family members IKZF1 (Ikaros) and IKZF3 (Aiolos) that are the lymphoid transcription factors crucial in myeloma cell survival ,117,118.Similar to the IMiD pharmacology, the arylsulfonamide indisulam and its analogs E7820 and tasisulam function via a molecular glue mechanism to drive the degradation of the essential RNA-binding protein RBM39 and the closely related splicing factor RBM23 (because of the high sequence conversation between their RBM domains involved in indisulam-induced molecular recognition) by chemically reprogramming the substrate DCAF5 ,126,127.Despite the therapeutic efficacy of drug-induced destruction of transcription factors and other cancer targets, a subset of proteins remain resistant to targeted degradation using existing approaches. An alternative mechanism has been described in which a small molecule induced the highly specific, reversible polymerization of a POI, followed by its sequestration into cellular foci and subsequent depletion . The smaAltogether, molecular glue degraders open highly appealing avenues for the development of antitumor therapeutic agents and synthetic biology.Specific and non-genetic inhibitors of apoptosis protein (IAP)-dependent protein erasers (SNIPERs) are chimeric molecules designed to induce IAP-mediated ubiquitination and proteasomal degradation of POI. They recruit the IAP family of RING-type E3 ubiquitin ligases—cIAP1, cIAP2 and XIAP with antiapoptotic properties. Cancer cells often overexpress IAPs to evade apoptosis with a concomitant increase in resistance to cancer therapy, making these proteins attractive drug targets. The chemical structure of a SNIPER consists of a selective IAP antagonist such as bestatin, MV1 and LCL161, a PEG linker and a peptide- or small-molecule-based POI specific component ,134,135.The capacity of the dominating TPD platforms such as PROTAC is limited to soluble intracellular proteins because of their dependence on the UPS. Harnessing an alternative intracellular degradation mechanism such as macroautophagy may broaden the target spectrum including aggregated proteins, non-protein biomolecules, and organelles. The first autophagy-mediated degraders are chimeric molecules called AUTACs (Autophagy Targeting Chimeras), that consist of a S-guanylation-inspired degradation tag and a specific binder of an intracellular target of interest ,138 Figa. Of notA complementary approach was proposed by autophagy tethering compounds (ATTECs) to target non-protein biomolecules or relevant organelles ,140. TheAnother chemical platform called AUTOphagy TArgeting Chimera (AUTOTAC) employs bifunctional molecules composed of target-binding ligands linked to autophagy-targeting ligands . AUTOTACUnlike AUTAC/ATTEC/AUTOTAC which utilize macroautophagy, CMA-based degraders harness chaperone-mediated autophagy (CMA) for protein degradation . These mThe cytosolic localization of the UPS and targetable autophagy machinery restricts these approaches to POI with cytosolic domains and requires degraders to be cell permeable. To expand the scope of TPD to extracellular and membrane associated proteins that make up to 40% of all protein-encoding genes and involve growth factors, cytokines and other key agents in cancer and other diseases, a novel platform called LYTAC (LYsosome TArgeting Chimeras) has been developed a. These In a similar attempt, Caianiello et al. developed a class of modular, bifunctional synthetic molecules termed MoDE-As (molecular degraders of extracellular proteins through the ASGPR) to drive the degradation of extracellular proteins . MoDE-A Recently, Han and colleagues constructed bispecific aptamer chimeras (BIAC) for lysosomal degradation of targeted membrane-associated proteins such as mesenchymal epithelial transition (Met) receptor and membrane receptor tyrosine protein kinase-like 7 (PTK-7), both known therapeutic cancer targets  Figure c.Wells and colleagues reported the development of an antibody-based PROTAC (AbTAC) that recruits membrane-bound E3 ligase RNF43 for the depletion of the cell-surface immune checkpoint protein programmed death-ligand 1 (PD-L1)  Figure d. The evAnother new strategy to combat cancer has been proposed through RIBOnuclease TArgeting Chimeras (RIBOTACs), which use RNA-targeting small molecules and RNase L, an otherwise latent ribonuclease, to accomplish the degradation of intracellular oncogenic RNAs . The firThe first wave of protein degraders has focused on the oncology fields. The target proteins can be categorized into the following groups: (i) those involved in cancer cell proliferation; (ii) in apoptosis; (iii) in angiogenesis; (iv) in immune evasion or inflammation; and (v) in cancer invasion and metastasis  50,51,1,157. AmoThe androgen receptor (AR) is a key driver of castration-resistant prostate cancer during the transition from a localized to a metastatic disease. Targeting the AR signaling axis with abiraterone, enzalutamide, darolutamide and apalutamide has demonstrated an overall survival benefit in the castration-sensitive state . HoweverL702H mutation and AR-V7 variance [T878 and ARH875 mutations [T878 and/or ARH875 mutations; (ii) ARL702H mutation or AR-V7; and (iii) wild-type AR or other AR alterations. A fourth subgroup involved patients based with a clinical history of less prior treatment strategy: ≤1 therapy for mCRPC, 1 novel hormonal agent, and no chemotherapy [T878A/S and/or ARH875Y mutations. Of 7 RECIST -evaluable patients with ART878A/S and/or ARH875Y mutations, 6 exhibited tumor shrinkage. Among the most common treatment-related adverse events were nausea, fatigue, vomiting, decreased appetite, diarrhea and alopecia; none were grade ≥4 (NCT03888612). In a parallel study, the combination of ARV-110 with abiraterone will be assessed in patients with metastatic prostate cancer with PSA increase on abiraterone (NCT05177042). ARV-110 oral tablets in combination with abiraterone and a corticosteroid will be administered daily in 28-day cycles. ARV-766 is another oral AR PROTAC with a different profile than ARV-110 [ARV-110  is the first-in-class PROTAC against AR. The treatment led to near-complete clearance of AR and suppressed AR-associated gene expression in vivo. ARV-110 inhibited the synthesis of prostate-specific antigen (PSA) and AR-dependent cancer cell proliferation by inducing apoptotic cell death. It also exhibited activity in enzalutamide refractory/resistant prostate cancer xenograft models with AR amplification and mutations with the exception of ARvariance ,160,161.utations . In phasotherapy . ARV-110 ARV-110 . A phase+). While approved treatments have achieved great success in this patient population, many ER+ breast cancers develop resistance to therapy [+ metastatic breast cancer following anti-estrogen therapy. Even though fulvestrant has verified the importance of ER degradation as a therapeutic intervention, up to 50% of ER can remain when compared to baseline levels after six months of treatment. Several ER-directed PROTAC degraders are being tested in preclinical and/or clinical settings [+ metastatic breast cancer. In preclinical studies, ARV-471 robustly degraded ER in ER+ breast cancer cell lines, suppressed expression of ER-target genes  and inhibited cell proliferation of ER-dependent cell lines  [Around 80% of all newly diagnosed cases of breast cancer are estrogen receptor positive  locally advanced or metastatic breast cancer to investigate the safety, tolerability and pharmacokinetics of ARV-471 (NCT05463952). The drug will be administered orally once daily with food in continuous dosing over 28-day cycles. A phase II neoadjuvant study will evaluate ARV-471 or anastrozole, a nonsteroidal aromatase inhibitor, in post-menopausal women with ER+/HER2- localized breast cancer amenable to surgical resection to address the biological activity of respective compound (NCT05549505). ARV-471 will be administered once daily and 1 mg anastrozole will be given once daily until surgery, which will take place approximately 5.5 months after starting treatment. The safety, tolerability and clinical activity of ARV-471 alone or in combination with palbociclib are addressed in the ongoing dose escalation and cohort expansion study in patients with ER+/HER2- advanced or metastatic breast cancer following prior chemotherapy or hormonal therapy (NCT04072952). The phase I part utilized traditional 3 + 3 dose escalation with ARV-471 oral tablets once daily for 28-day cycles in 21 heavily pre-treated patients with poorer prognosis. 48% of patients had visceral metastatic disease often in the liver and lung. All patients received previous CDK4/6 inhibitors; 71% of patients received fulvestrant; 38% of patients had chemotherapy; and 24% of patients had other selective ER degraders in the clinical trial setting. They had a median of 4 previous lines of therapy for their advanced or metastatic breast cancer. ARV-471 was well tolerated, the highest dose being 360 mg, and no grade 3/4 adverse events were observed. In paired tumor biopsy samples before and after treatment with ARV-471, mean ER degradation was significantly higher than with fulvestrant and has been observed in patients with either wild-type ER or mutant ERY537S, ERY537N, and ERD538G in their tumors. A further study is recruiting to assess the combination of ARV-471 and everolimus, an mTOR inhibitor, in patients with advanced or metastatic ER+/HER2- breast cancer who have received a prior CDK4/6 inhibitor and endocrine therapy in the advanced/metastatic setting (NCT05501769). Post-menopausal women or pre-/peri-menopausal women with ovarian suppression will receive ARV-471 oral tablets in combination with everolimus administered daily in 28-day cycles. A new sub-study aims to evaluate the safety and effects of ARV-471 when given together with the CDK4/6 inhibitor abemaciclib for the treatment of advanced or metastatic breast cancer (NCT05548127). The participants are enrolled based on the following criteria: (i) tumor is advanced or metastatic and cannot be fully treated by surgery or radiation therapy; (ii) tumor is sensitive to hormonal therapy; and (iii) tumor is nonresponsive to previous treatments. ARV-471 will be administered orally once daily, while abemaciclib will be given orally twice a day. The medication will continue until the tumors are not responding or side effects become too severe.Several clinical trials with ARV-471 are underway. A phase I, open-label study will recruit Japanese patients with ER+/HER2− locally advanced or metastatic breast cancer . These studies either involve post-menopausal patients with life expectancy ≥3 months (NCT05080842) or those with ≥1 prior endocrine therapy regimen, concomitant use of CDK4/6 inhibitor being allowed (NCT05489679).Other phase I clinical trials focus on AC682, an orally available ER degrader given as a single agent in 28-day cycles to address safety/tolerability, pharmacokinetics and antineoplastic activity in patients with ERBruton’s tyrosine kinase (BTK) plays a crucial role in B cell development, differentiation and signaling. BTK is closely associated with chronic B-cell receptor (BCR) activation, and is critical for the survival of B-cell neoplasms. Ibrutinib, a first-in-class covalent BTK inhibitor, has been approved for the treatment of several types of Non-Hodgkin’s lymphoma, specifically relapsed/refractory mantle cell lymphoma, chronic lymphocytic leukemia and Waldenström macroglobulinaemia. However, resistance eventually develops, highlighting the dire need for other strategies.NX-2127 is not only a BTK PROTAC but also a molecular glue for IKZF1/3 degradation due to its cereblon binding activity to recruit ubiquitin ligase complex for BTK degradation . The comBRD4, a member of the bromo- and extra-terminal (BET) family of proteins, recruits transcriptional regulatory complexes to acetylated chromatin, thereby controlling specific networks of genes critical for proliferation and cell cycle progression . AlteratBromodomain-containing protein 9 (BRD9), a specific component of the non-canonical mammalian SWI/SNF chromatin remodelling complex, is essential to maintain the transformed phenotype of acute myeloid leukemia and the growth of synovial sarcoma ,193. ThiSTAT3, a protein that has been historically undruggable, is a transcriptional regulator allied with numerous cancers and other inflammatory and autoimmune disorders. Under steady state conditions, the STAT3 protein is activated by cytokines and growth factors, thereby triggering transcriptional regulation of many cellular functions, while under stress conditions such as cancer, STAT3 activity becomes dysregulated, resulting in persistent activation of STAT3, which is associated with poor prognosis. Selective degraders targeting STAT3 thus have the potential to provide an intriguing solution to address multiple STAT3-dependent pathologies. KT-333 is a first-in-class PROTAC of STAT3. The FDA has granted orphan drug designation to KT-333 for the treatment of peripheral T-cell lymphoma, a disease entity with dysregulated STAT3 for which no approved therapies are available . An openL inhibitors, such as orally bioavailable ABT263, as safe and effective anticancer agents. DT2216 is a selective BCL-xL degrader utilizing ABT263 and a VHL E3 ligase binder [+ T cells [+ T cell-depleted mice. Thus, BCL-xL destruction represents a novel avenue for cancer immunotherapy. Based on these promising preclinical findings, DT2216 is under clinical evaluation in a dose escalation and cohort expansion study for its safety, tolerability, and activity in patients with relapse and refractory solid tumors and hematologic malignancies when administered intravenously in 28-day cycles (NCT04886622).B-cell lymphoma extra large (BCL-xL) is a well-validated cancer target. However, the on-target and dose-limiting thrombocytopenia limits the use of BCL-Xe binder . DT2216 e binder . DT2216  T cells . These aV600X, which accounts for approximately 50,000 cancer diagnoses annually, to offer more durable responses. The bifunctional CFT1946 received “the study may proceed letter” from the FDA to initiate a phase I/II clinical study in patients with BRAFV600 mutant solid cancers including lung, colorectal and melanoma [V600E-driven disease and in the escape mutant BRAFV600E/NRASQ61K-driven model [V600E degradation by CFT1946 caused loss of MAPK signaling superior to inhibition alone and attenuated viability of BRAFV600E cells but not wild-type BRAF cells. Another PROTAC which has been approved for the investigational new drug (IND) application is the world’s first-in-class NTRK degrader  for the treatment of advanced solid tumors [L858R in non-small cell lung cancer [CFT1946 is an orally bioavailable mutant selective degrader of BRAFmelanoma . The trien model . In precd tumors . One PROg cancer . Precling cancer . KT-253 g cancer . As wildIMiDs are among the standard of care for treatment of multiple myeloma and have shown activity in some non-Hodgkin’s lymphoma subtypes. Given that many patients treated with these agents often develop disease progression, an unmet need remains. New generation IMiDs have been designed for better clinical efficacy . AvadomiInterleukin-1 Receptor-Associated Kinase 4 (IRAK4) is a key mediator of innate immunity. It is found hyperactivated in a variety of autoimmune diseases. Multiple small-molecule kinase inhibitors of IRAK4 have been developed to block its kinase activity in autoimmune, inflammatory and oncological diseases, with the most advanced being in phase II clinical trials. However, some reports have indicated a nonkinase function for IRAK4 in several cell types, pointing out the need of alternative therapeutic interventions . KT-413 Malignant tumors must evade the immune system for continuous growing, a hurdle some malignancies may overcome by attracting immune-suppressive regulatory T cells (Tregs). The zinc finger transcription factor IKZF2 (Helios) is critical for the activity and stability of Tregs; its deficiency enhances immune responses to tumors in vivo, indicating IKZF2 may be an appealing target for cancer immunotherapy. DKY709 is the first CRBN modulator agent targeting IKZF2/4 with no activity towards IKZF1/3. Upon exposure to DKY709, human Tregs showed reduced suppressive activity . Oral trOther CRBN modulators including CC-885, CC-90009, ZXH-1-161 and oral bioavailable candidates SJ6986 and MRT-2359 exhibit therapeutic potential for GSPT1 degradation ,219,220.Over the last two decades, the field of targeted protein degradation has evolved successfully, but almost all of this progress has been catalyzed through PROTACs alone. PROTACs have demonstrated excellent preclinical activities in multiple types of cancers by robust and selective depletion of their targets. Potent tumoricidal activities could also be validated in some in vivo models that were resistant to the counterpart small-molecule inhibitors of PROTACs. Despite their success, there are still some challenges concerning optimal druggability and evaluation of biological activity. Artificial intelligence and virtual drug screening platforms may improve identification of highly specific POI/E3 ligands. Molecular dynamic simulations, X-Ray crystallography and cryo-electron microscopy may be effective techniques for determining better ternary complex structures of PROTAC with POI and E3 ligase . Drug-li+/HER2- breast cancer, respectively. Although degrader drugs that are under clinical investigation (PROTAC and molecular glues, see Targeted protein degradation is one of the fastest growing fields. Within a relatively short time, the platform is already impacting medicine as multiple therapeutic degraders have entered the stage of practical application from conceptual frameworks . ClinicaResults of ongoing clinical trials are expected to establish the clinical significance of degrader drugs in oncology and other disease entities. Evidence of long-term effects or potent toxicity is still missing. Future clinical investigation will put more emphasis on the assessment of the clinical efficacy and adverse events in larger cohorts, exploration of potential resistance mechanisms, defining biomarkers for optimal selection of patients and exploration of potential synergy with other cancer therapeutics. In fact, early clinical investigations suggest that combination of chemical degraders either with targeted inhibitors or with chemotherapy/other agents may represent a robust alternative path for cancer therapy: ARV-471 produced a synergistic tumoricidal effect when combined with the CDK4/6 kinase inhibitor.Potentials of targeted degradation applications go well beyond what was thought to be possible. PROTACs could be used as immune-enhancing agents. In combination with immune checkpoint inhibitors, they could augment the ability of the immune system to recognize and erase tumors. Degrader systems against immune checkpoint proteins and those that modulate T-cell receptor function have already been developed ,226,227.The incorporation of new elements into PROTAC technologies may help taming the undruggable drug targets and thus broaden the landscape of targeted degradation. This is highlighted by recent reports of utilizing RNA and DNA as respective recruiting elements to eliminate unwanted targets for cancer prevention and treatment ,100,101.


Alcohol use predicts short- and long-term memory impairment. However, it is unclear how alcohol influences subjective memory, which refers to perceptions of one’s own memory functioning. This study examined associations between daily alcohol use and memory lapses  and evaluated perceived impact of memory lapses on daily life. Participants  were non-abstaining adults from the Midlife in the United States (MIDUS) study who participated in the 8-day daily diary project. Survey items included number of drinks and 9 memory lapse items (1=yes). After each reported memory lapse, participants indicated memory lapse irritation (range=1-10) and interference (range=1-10). Covariates included age, gender, race, education, and cognitive functioning. Multilevel logistic regression analysis revealed that within-person alcohol use , but not between-person alcohol use (p>0.05), was associated with greater odds of reporting any memory lapses. When assessing retrospective and prospective lapses separately, only the prediction of prospective lapses from between-person alcohol use was significant . Finally, multilevel linear regression analysis indicated that neither within-person nor between-person alcohol use was predictive of reported irritation or interference from memory lapses (p>0.05). Study findings revealed between- and within-person associations between alcohol use and subjective memory. At the daily-level, individuals were more likely to notice memory lapses on days they drank more than usual. At the person-level, people who drank more relative to others were more likely to report prospective memory lapses. Future studies should assess how associations between alcohol use and subjective memory relate to objective cognitive functioning outcomes.


Anti-N-methyl-D-aspartate receptor (anti-NMDA-R) encephalitis is well-characterised autoimmune encephalitis with prominent psychiatric manifestations, neurological manifestations like speech dysfunction, seizures, dyskinesias and other movement abnormalities, decreased level of consciousness and autonomic instability. This disorder affects primarily children and adults up to 45 years. Females are 4 times more common than males and may have association with ovarian teratoma.To identify anti-NMDA receptor encephalitis based on clinical features, facilitate early screening and relevant investigations to prevent delay in treatment.A case study of 36 year old female presented with clinical manifestations of autoimmune encephalitis syndrome.Diagnosis confirmed by presence of NMDA receptor antibodies in serum and cerebrospinal fluid.Early recognition of clinical features of Anti-NMDA receptor encephalitis and early initiation of treatment has shown to improve outcomes, speed recovery and reduce the risk of relapses.No significant relationships.


Esophageal cancers feature distinct manifestations between and within patients which complicate precision diagnosis, prognosis, and patient care. New genomic and epigenomic research uncovers novel mechanisms underlying both inter- and intra-tumoral heterogeneity in esophageal cancer, with significant biological and translational implications. Inter- and intra-tumoral heterogeneity are both recognized to strongly affect cancer diagnosis, prognosis and clinical management, thereby contributing to treatment failure and poor clinical outcome.Esophageal squamous cell carcinoma (ESCC) is a common and lethal malignancy worldwide. A number of tumor sequencing studies have hitherto established the genomic landscape of ESCC; however, these efforts have led to neither genome-guided targeted therapies nor molecular-based diagnostic or prognostic biomarkers. Instead, genomic analyses have underscored highly heterogeneous genomes of ESCC, manifesting in both inter- and intra-tumoral forms2. The high inter-tumoral heterogeneity and long-tailed mutation list also suggest that most of genomic drivers and druggable mutations are present only in a small fraction of ESCC tumors, an obvious obstacle for implementing the panel-sequencing approach for patient stratification and selection. Intra-tumoral heterogeneity fuels cancer evolution and diversification, causing immune evasion, drug resistance and disease recurrence. Moreover, intra-tumoral heterogeneity often leads to sampling bias, hindering precision diagnosis and personalized medicine.The ESCC genome exhibits high inter-tumoral heterogeneity and yet moderate-to-low mutational burden, posing a key challenge for comprehensively identifying genomic drivers and actionable mutations. In fact, based on the background mutational rate of ESCC, saturation analyses estimated that identification of driver mutations in 2–3% of the patients requires sequencing of 1500-2000 tumor genomes3. Undoubtedly, an urgent need exists to characterize further ESCC genomes for the development of personalized treatment and oncological biomarkers. This requires not only sequencing a large number of patients to overcome its strong inter-tumoral heterogeneity, but also characterizing in detail the clonal makeup and evolution to decipher ESCC intra-tumoral heterogeneity. New research5 published recently in Nature Communications have specifically taken on these two challenges, by performing genomic and epigenomic analyses with significant breadth and depth, and shed novel insights into ESCC tumor heterogeneity with translational implications.Clinically, ESCC tumors are often resistant to cytotoxic chemoradiotherapy; ESCC patients suffer from dismal outcomes, having a 5-year survival rate lower than 20%5 curated and integrated ESCC sequencing data from 1930 patients across 33 studies, representing one of the largest compendia of paired tumor-normal sequences. To ensure data quality and minimize batch effects, the authors paid extra attention and made efforts during quality control, data processing, homogenization and verification. The large number of sequencing data revealed several new significantly mutated genes in ESCC, including PPFIA2, TGFBR2, ZBBX, ATP13A5 and IRF2BPL. In addition, novel recurrent frameshift deletions in TGFBR2 and IRF2BPL were identified, indicative of their tumor suppressing functions. The collection of clinicopathological and epidemiological information of most sequenced patients, coupled with high statistic power, enabled robust clinical associations. For example, NOTCH1 mutations were prominently associated with the increased diagnostic age of ESCC patients, consistent with previous findings of NOTCH1 genomic alterations in normal esophageal tissues of elderly individuals6. Both log rank test and multivariable adjusted Cox analysis revealed gene mutations associated with inferior prognosis in both early- and late-stage patients. On the other hand, certain gene mutations were associated with stage-specific survival. For example, NFE2L2 mutations were specifically correlated with poor survival of late-stage patients. This association is likely explained by prior findings that NFE2L2 mutations conferred chemotherapy and radiation resistance in ESCC cells7.To address the inter-tumoral heterogeneity, Li et al.in NFE2L2, CSMD1, CREBBP, KALRN, PRUNE2, NRXN1, AKAP9 and FREM2. In early-stage patients, 1-2 and >2 positive mutations were respectively associated with 1.78 and 2.26 of hazard ratio (HR), indicating substantial predictive power. A similar result was found in late-stage patients. The prognostic property was validated in multiple independent datasets upon stage adjusted HR in Cox regression, confirming the mutational score as a robust prognostic model in ESCC. Further facilitating its potential clinical use, the eight-gene panel was able to cover a considerable proportion of ESCC patients, with 29.1% early- and 27.8% late-stage patients having at least one mutation. This detection rate substantially overcomes the challenge of inter-tumoral heterogeneity of ESCC patients.Nevertheless, most of the mutations with prognostic values occurred in less than 5% ESCC patients, limiting their potential clinical application as biomarkers individually. The authors thus built a statistical model to establish a gene-panel-based mutational score for prognosis prediction. Upon balancing the positive rate and the complexity of the model, an eight-gene mutational score was established, defined as the sum of nonsynonymous mutations 4, Cui and colleagues performed one of the largest analyses of intra-tumoral heterogeneity of ESCC, characterizing 186 primary samples from 36 patients. Multi-omic sequencing, including whole exome sequencing (WES), reduced representation bisulfite sequencing (RRBS) and RNA-seq were performed on the majority of samples. This comprehensive strategy enables in-depth analyses of genomic, epigenomic and transcriptomic intra-tumoral heterogeneity and the interplay between these mechanisms during tumor evolution, addressing several key questions.In the other studyPerhaps one of the most intuitive questions is what are the molecular mechanisms driving intra-tumoral heterogeneity at different molecular levels. The study provided compelling evidence showing that chromosomal instability is the most significant force promoting both genomic and epigenomic intra-tumoral heterogeneity. Indeed, genome doubling was associated with subclonal copy-number alterations. Epigenomically, somatic copy number altered regions showed a higher degree of epigenetic entropy and DNA methylation variability. At the transcriptomic level, genes with subclonal copy number gains had higher mRNA expression and the reverse was also true. Additionally, variable promoter methylation levels also contribute to the intra-tumoral expression heterogeneity.8. This is what one would expect from those loci where DNA methylation reflects a passive mitotic clock. However, the evolution trajectories of somatic mutations and DNA methylation were largely discordant in liver cancer9. This would be expected for methylation loci that are associated with active gene regulatory elements rather than a passive mitotic clock. Here4, both concordant and discordant patterns of phylogenetic trees were also noted in this large collection of intra-tumoral sequencing data. Clarifying these disparities thus warrants future investigation of the associations between mitotic turnover vs. gene regulation.It is unclear if genomic and epigenomic changes converge or diverge during tumor evolution and why. Different patterns have been reported regarding the phylogeny relationship between somatic mutations and DNA methylation in different cancer types. For example, in an earlier study of ESCC, the phyloepigenetic tree of DNA methylation changes followed exactly the tree of passenger mutations in the genome4 first deconvoluted RNA-seq data, revealing diverse levels of immune infiltrates across different spatial regions within the same tumor, which was further validated by orthogonal multiplex immunostaining. The intra-tumoral variability of somatic mutations was associated with immune infiltration diversity, echoing a previous finding of a strong correlation between the repertoire of T cell receptors  and genomic intra-tumoral heterogeneity in ESCC10. Neoantigens were then predicted bioinformatically, and it was noted that intra-tumoral heterogeneity of neoantigens was stronger in tumors with more diverse immune infiltration, indicating the specific selective pressure on neoantigens by tumor-infiltrating lymphocytes. In probing the mechanisms underlying immune escape, the authors identified that out of 36 patients, nine harbored clonal loss of heterozygosity (LOH) of HLA and two with subclonal LOH, suggesting that LOH of HLA is an early and prevalent genetic mechanism behind immune evasion. In addition to copy number changes, mRNA expression of HLA was also interrogated. Importantly, intra-tumoral variability of infiltrating CD8+ T cells and NK cells were significantly correlated with the intra-tumoral expression heterogeneity of HLA-A, HLA-B, and B2M, suggesting that the intra-tumoral expression heterogeneity of HLA genes is driven by and adaptive to diverse immune cell attacks.Another important question with translational significance is how the cross-talk between tumor cells and immune microenvironment influences cancer evolution. Relatedly, it is unclear what are the mechanisms promoting clonal and subclonal immune evasion. The authors5 have uncovered novel mutational features exhibiting robust associations with key clinical parameters, revealed molecular mechanisms driving clonal diversification, and characterized the interplay between immune selection pressure and tumor evolution. Moving forward, new single-cell genomic and epigenomic technologies are revolutionizing the research on tumor heterogeneity, and recent single-cell RNA-seq studies13 have begun to highlight the extensive diversity of gene expression programs among ESCC cancer cells. For example, prominent inter- and intra-tumoral expression heterogeneity at both individual gene level and the pathway level were conspicuous in these single-cell datasets13. Moreover, certain gene expression programs from subsets of tumor cells were associated with the variability of immune infiltration, indicating the cross-talk between diverse clusters of malignant cells and immune populations13. The future development of methodologies to profile somatic mutations, chromatin accessibility and DNA methylation from the same cells will provide an exceptional opportunity to investigate the relationship between phylogenetic and phyloepigenetic architectures during cancer evolution. Moreover, the status quo of tumor heterogeneity research remains static and descriptive, with a clear gap between tumor sequencing and functional annotation of cancer genome and epigenome. One kind of models that would allow for such functional interrogation in a versatile and robust manner are patient-derived tumor organoid cultures. In fact, organoid modeling has been successfully applied to investigate both inter-14 and intra-tumor15 heterogeneity in several cancer types. The exciting new technologies, combined with advanced theoretical understanding of tumor heterogeneity, will significantly accelerate the development of precision diagnosis, prognosis, and personalized care of ESCC patients.In summary, by performing large-scale genomic investigation, combined with in-depth computational and statistical analyses, these new studiesFurther information on research design is available in the Reporting Summary


Hypoxic-ischemic encephalopathy is brain injury resulting from the loss of oxygen and blood supply around the time of birth. It is associated with a high risk of death or disability. The only approved treatment is therapeutic hypothermia. Therapeutic hypothermia has consistently been shown to significantly reduce the risk of death and disability in infants with hypoxic-ischemic encephalopathy. However, approximately 29% of infants treated with therapeutic hypothermia still develop disability. Recent preclinical and clinical studies have shown that there is still persistent neuroinflammation even after treating with therapeutic hypothermia, which may contribute to the deficits seen in infants despite treatment. This suggests that potentially targeting this persistent neuroinflammation would have an additive benefit in addition to therapeutic hypothermia. A potential additive treatment is Exendin-4, which is a glucagon-like peptide 1 receptor agonist. Preclinical data from various in vitro and in vivo disease models have shown that Exendin-4 has anti-inflammatory, mitochondrial protective, anti-apoptotic, anti-oxidative and neurotrophic effects. Although preclinical studies of the effect of Exendin-4 in perinatal hypoxic-ischemic brain injury are limited, a seminal study in neonatal mice showed that Exendin-4 had promising neuroprotective effects. Further studies on Exendin-4 neuroprotection for perinatal hypoxic-ischemic brain injury, including in large animal translational models are warranted to better understand its safety, window of opportunity and effectiveness as an adjunct with therapeutic hypothermia.  Moderate to severe hypoxic ischemic-encephalopathy (HIE) affects 1–3 in 1000 live term born infants in high income countries and is associated with a high risk of death or lifelong disability ,3. AlthoAdditionally, the recent hypothermia for moderate or severe neonatal encephalopathy in low-middle income countries (HELIX) trial showed that therapeutic hypothermia did not reduce the combined outcome of death and disability, but increased the risk of death . HoweverThere is thus considerable commitment to finding additive treatments to therapeutic hypothermia. Pre-clinical studies have shown that there is persisting neuroinflammation even after treatment with hypothermia ,11. For Perinatal hypoxic-ischemic brain injury evolves over time and is classified into four phases  13]. Th. Th13]. The inflammatory response after hypoxia-ischemia can be initiated within minutes after the insult, and can persist for weeks to months . NeuroinThis inflammatory response involves the resident immune cells in the brain including microglia and astrocytes, and the infiltrating peripheral immune cells such as neutrophils, mast cells and macrophages. These cells release pro-inflammatory factors such as cytokines, chemokines, nitric oxide (NO), reactive oxygen species (ROS), excitatory amino acids and death receptor ligands ,29,30. TMicroglia are the resident immune cells of the brain and they play an important role in immune surveillance under normal conditions . RestingActivated microglia have various roles, however, the classification of these functions remain controversial . The claIn postnatal day (P)7 rats, there was a significant increase in CD45/CD11b positive cells in injured areas assessed at 24 h after transient middle cerebral artery occlusion (MCAO). Interestingly, these cells were predominantly microglia and not infiltrated blood monocytes . SimilarThe functions of activated microglia appear to evolve over time after hypoxia-ischemia. The activation state of microglia and infiltrated macrophages has been characterized in P9 mice after common carotid artery occlusion followed by hypoxia . This stAstrocytes are another cell type that can contribute to the inflammatory response after perinatal brain injury. During reactive astrogliosis, astrocytes become hypertrophic and upregulate expression of glial fibrillary acidic protein (GFAP) and potentially form a glial scar around a focal injury . A potenAlthough astrocyte responses are well-characterized in response to ischemic brain injury in the adult brain, the role of reactive astrogliosis in the developing brain is less well understood . DiffereThe neuroprotective effect of therapeutic hypothermia is mediated through a range of mechanisms beyond merely the suppression of metabolism, including its anti-apoptotic, anti-excitotoxic, anticonvulsant and anti-inflammatory effects, as reviewed in . This seA key mechanism of therapeutic hypothermia is its anti-inflammatory effect. In an in vitro study, hypothermia started before, during and immediately after exposure to hypoxia reduced microglial activation measured by reduced inducible nitric oxide synthase (iNOS) production in both BV-2 microglial cell line and mouse primary microglia cultures . HypotheRecent data have shown that numbers of microglia in the parasagittal cortex and in the intragyral white matter of the parasagittal gyrus were both inversely correlated with EEG power at day 7 after global cerebral ischemia in near-term fetal sheep . That isEx-4 is a 39 amino acid peptide found in the saliva of the Gila Monster (Heloderma suspectum) . Ex-4 isA seminal study of the effect of Ex-4 (4 doses 0.5 µg/kg i.p.) in P7 mice showed that there was a reduction in infarction size, cell death and inflammation associated with Ex-4 treatment started at 2 h after common carotid ligation and hypoxia . When Ex+ regulatory T cells [Ex-4 has been shown to have profound anti-inflammatory effects that may contribute to neuroprotection. Ex-4 administration in neonatal mice born to dams exposed to systemic inflammation induced by lipopolysaccharide (LPS) was associated with increased numbers of IL-10 expressing neutrophils, and suppressed expansion of CD8 T cells . The num T cells   in lung and large intestine tissues in neonatal mice born to dams exposed to systemic inflammation induced by LPS [Ex-4 has also been shown to modulate microglial polarization to be more “anti-inflammatory” ,78,79. Fd by LPS . In humad by LPS . This wad by LPS . FurtherPre-clinical studies have shown that Ex-4 can modulate inflammation through many different inflammatory pathways. For example, in peripheral blood mononuclear cells collected from patients with diabetes, treatment with Ex-4 reduced the secretion of pro-inflammatory cytokines TNF, IL-1β and IL-6, mediated through the suppression of the mitogen-activated protein kinase (MAPK) signaling pathway . In an aEx-4 has mitochondrial protective effects that likely contribute to neuroprotection. Ex-4 improved mitochondrial function, integrity and respiratory control rate in rodent models of Alzheimer’s disease ,84. TherEx-4 has consistently reduced apoptotic cell death in models of perinatal hypoxic-ischemic brain injury, stroke and spinal cord injury ,69,76,85Another mechanism contributing to the neuroprotective effect of Ex-4 may be its anti-oxidative effects. For example, Ex-4  reduced oxidative stress, shown by a reduction in the number of 8-Hydroxy deoxyguanosine (oxidative DNA damage product) and 4-hydroxy 2-hexenal (lipid peroxidation product) positive cells in adult mice after MCAO . AdditioEx-4 has also been shown to have neurotrophic effects. For example, Ex-4 administration  to diabetic mice was associated with an increase in the expression of neurotrophin BDNF and CREB a transcription factor regulating BDNF expression in the hippocampus . In turnWe have discussed the broad range of effects of Ex-4 and its potential for treating perinatal hypoxic-ischemic brain injury. The next pragmatic question is when is the most beneficial time to administer Ex-4? In the Rocha-Ferreira study, Ex-4 was administered at the same time as the start of hypothermia (10 min after hypoxia-ischemia), which was associated with an improved macroscopic tissue score compared with hypothermia alone. Therefore, this study suggests that concurrent treatment is beneficial. However, given there is persistent inflammation following therapeutic hypothermia , this raEx-4 is currently only approved for use in adults for treating type II diabetes. Therefore, if sufficient pre-clinical data show that Ex-4 is neuroprotective, the safety profile of Ex-4 in infants will have to be carefully established. Ex-4 treatment did not alter blood glucose levels in neonatal mice, compared with control animals . This isThere are other FDA approved GLP-1R agonists available, which are gaining increasing interest for neuroprotection. Liraglutide is a modified form of the human GLP-1 protein, sharing 97% homology with GLP-1, but with an extended elimination half-life compared with GLP-1 . To our There is now considerable evidence of prolonged neuroinflammation after hypoxia-ischemia. This continuing inflammation may, at least in part, contribute to the functional and histological deficits seen clinically and experimentally even after treatment with hypothermia. This suggests that neuroinflammation may be an important target to further improve treatment of HIE. Ex-4 is a promising drug that has potent anti-inflammatory effects and, after early intervention, has shown neuroprotective effects in neonatal mice exposed to HI . Excitin


The incidence of obesity and associated metabolic diseases is increasing globally, adversely affecting human health. Dietary fats, especially triglycerides, are an important source of energy for the body, and the intestine absorbs lipids through a series of orderly and complex steps. A long-term high-fat diet leads to intestinal dysfunction, inducing obesity and metabolic disorders. Therefore, regulating dietary triglycerides absorption is a promising therapeutic strategy. In this review, we will discuss diverse aspects of the dietary triglycerides hydrolysis, fatty acid uptake, triglycerides resynthesis, chylomicron assembly, trafficking, and secretion processes in intestinal epithelial cells, as well as potential targets in this process that may influence dietary fat-induced obesity and metabolic diseases. We also mention the possible shortcomings and deficiencies in modulating dietary lipid absorption targets to provide a better understanding of their administrability as drugs in obesity and related metabolic disorders. Obesity is a growing global health crisis. Obesity leads to a number of comorbidities such as non-alcoholic fatty liver disease (NAFLD), diabetes, hyperlipidemia, cardiovascular disease, and cancer, causing an enormous health burden and harm life quality . To curbIncreased food energy consumption is the leading cause of obesity and metabolic diseases . TriglycThe intestine, which absorbs nutrients, is of principal importance in regulating dietary fat absorption. Nearly 95% of dietary TG can be absorbed and undergo digestion, uptake, resynthesis, and secretion into the circulation in chylomicron (CM) . Dietarysn-2-monoacylglycerol (2-MAG) and fatty acids (FAs). 2-MAG and long-chain FA (LCFA) are taken up by intestinal epithelial cells effectively  to yield ectively .in vitro experiments suggest that FAs transporters are partially involved in this process , fatty acid transport protein 4 (FATP4), and the plasma membrane-associated fatty acid-binding protein, are involved in the transportation process . Moreove process .LCFAs and 2-MAG traverse across the cytoplasm to reach the endoplasmic reticulum (ER) membrane in enterocytes. Cytoplasmic fatty acid-binding proteins such as intestinal FABP (I-FABP) and liver FABP (L-FABP) are involved in intracellular LCFAs transportation .The first step of TG resynthesis is adding the coenzyme A (CoA) group to the LCFAs, which is catalyzed by long-chain acyl-CoA synthetases (ACSL) . FATP4 hIn the ER membrane, two multi-enzymatic systems catalyze FA-CoA re-esterified into TG: the glycerol-3-phosphate pathway and the monoacylglycerol pathway. In enterocytes, TG resynthesis is mainly along the monoacylglycerol pathway, and nearly 80% of the resynthesized TG is produced from this pathway . Monoacyde novo FAs synthesis. CLDs synthesized from different sources may undergo different metabolic fate. FAs delivered from the apical side are preferably used for TG synthesis, while the FAs delivered from the basolateral side are used for phospholipid synthesis and preferential oxidation  in a highly dynamic state . In entexidation .via pre-CM transport vesicles (PCTVs) for further maturation. In Golgi, pre-CM is further modified by addition of ApoA-I and glycosylating ApoB-48 to form mature CM [reviewed in  facilitates lipidation of apolipoprotein B-48 (ApoB-48) to form the pre-CM. The pre-CMs is then secreted from the ER and transported to the Golgi iewed in ]. Matureiewed in .The amount of dietary TG intake is a decisive factor in obesity and metabolic diseases. The human body has a good absorption and clearance capacity of dietary TG. A single intake of 15 g of dietary TG did not affect postprandial lipids and lipoprotein in healthy adults . Althougsn-1 and sn-3 sites of TG to form free FAs and 2-MAG  for 8 weeks . Accordiood flow .et al. demonstrated that reduced numbers of small intestinal intraepithelial lymphocytes and lamina propria lymphocytes could be found in mice fed HFD for only 1-day. HFD feeding for 3 weeks induced atrophy of gut-associated lymphoid tissue  and lipoprotein synthesis (ApoA-IV) gene expression  expansion of the size of CMs, ii) an increase in the number of CMs, and iii) a combination of these processes. In insulin-deficient rat model, the secretion rate of CMs is decreased, whereas the size of CMs is increased . To adap obesity . The inc obesity . CMs and obesity .−/−) have altered BA composition, impaired absorption of dietary TG, increased fecal TG levels, decreased body weight and improved glucose tolerance (−/− mice treated with 12α-hydroxylated BA taurocholic acid or CA restored the absorption of lipids (Bile acids (BAs) emulsify dietary lipids and facilitate the hydrolysis and absorption of fats in the proximal intestine. The composition of BAs can determine the absorption efficiency of dietary lipids. Mice lacking 12α-hydroxylase cytochrome P450 family 8 subfamily B member 1gene (Cyp8b1olerance . The 12αolerance . Cyp8b1−f lipids . Howeverf lipids  Table 1−/− have PL is responsible for the hydrolysis of 50%–70% of dietary lipids . PL inhiCd36 gene deficiency (Cd36−/−) mice showed a 50% reduction in FA uptake in proximal enterocytes  mice exhibited no modified fat absorption and had no protection against obesity during HFD feeding  mice showed protection against diet-induced obesity, partly due to the higher presence of steatorrhea  mice decreased circulating free FAs and low-density lipoproteins cholesterol levels, preventing HFD-induced obesity and rapid increase in low-density lipoproteins cholesterol  mice exhibited a leaner phenotype when fed a HFD than wild-type mice, whereas L-Fabp deletion (L-Fabp−/−) mice displayed higher body weight and body fat mass  mice exhibited decreased TG absorption, lower fat mass, lower blood TG and glucose levels, and increased insulin sensitivity without acute postnatal complications  exhibited resistance to HFD-induced obesity, although they consumed and absorbed normal amounts of dietary fat  increased energy expenditure in mice, improved leptin and insulin sensitivity, and improved resistance to diet-induced obesity and NAFLD  is a prominent phospholipid found in CLD membranes. Lysophosphatidylcholine acyltransferases3 (LPCAT3) are involved in the re-acylation of PC in the intestine . Intesti protein . Althougteatosis .Cideb (Cideb−/−) had lower levels of plasma TG, improved IR and protection against HFD-induced obesity and fatty liver  family proteins, which are enriched at CLD-CLD contact site, mediate the transfer of lipids from small CLDs to large CLDs . In the ty liver . In the pidation .Atgl (AtgliKO) increases CLDs in enterocytes but does not influence TG absorption, suggesting that CM secretion might be ATGL-independent , hormone-sensitive lipase, and monoacylglycerol lipase (MGL). First, ATGL hydrolyzes TG to FA and DAG. Intestinal-specific deletion of ependent . During ecretion . In compnditions . Intestiaffected . The synaffected . Consistteatosis . ATGL plteatosis .Hormone-sensitive lipase catalyzes the conversion of DAG to FA and MAG. Interestingly, mice with intestine-specific hormone-sensitive lipase deletion did not display impaired TG metabolism but had increased plasma cholesterol concentrations and cholesteryl ester accumulation in the small intestine, suggesting that hormone-sensitive lipase may play a more important role in maintaining cholesterol homeostasis .et al. demonstrated that the primary function of MGL was to convert dietary MGs to TG instead of CLD degradation in the intestine (Mgl overexpression (iMgl) mice exhibited unaltered dietary fat absorption. Body weight, hepatic and serum TG levels of iMgl mice were higher than those of wild-type mice after only 3 weeks of HFD feeding. This may be due to the decreased levels of 2-arachidonyl glycerol caused by Mgl overexpression, which led to the hyperphagic behavior and decreased energy expenditure  showed impaired CM assembly, increased accumulation of TG in enterocytes, and no CM particles in the secretory pathway. However, although MtpiKO mice exhibited decreased plasma and hepatic TG content, hepatic lipogenesis and low-density lipoproteins secretion increased, suggesting an unexpected interaction between intestinal and hepatic lipid metabolism  reproduce some chylomicron retention disease features. Sar1b−/− mice exhibited decreased ApoB and Mtp expression, contributing to the malabsorption of fat and decreased secretion of CM (+/+) can enhance intestinal fat absorption under HFD condition. Moreover, Sar1b +/+ mice showed phenotypes of dyslipidemia, obesity, and liver steatosis and were susceptible to IR  proteins. The binding of L-FABP to the ER initiates PCTV budding. activity . SAR1B iactivity . Mice wion of CM . Corresple to IR . In Cacoelevated .via VAMP7 can improve insulin sensitivity without affecting dietary fat absorption  mice, CM transport out of enterocytes was impaired, and massive CM secretory vesicles were accumulated in the enterocytes, as CM secretory vesicles from the Golgi failed to fuse with the basolateral membrane. Dennd5b−/− mice were resistant to western diet-induced obesity, exhibited improved plasma and hepatic lipid concentrations and reduced aortic atherosclerosis  regulates lipid entry into the lacteals. Plagl2 deletion mice develop severe emaciation due to the blockage of CMs from the lamina propria into the lacteals, and exhibit growth arrest and even die of starvation shortly after birth. Plagl2 null enterocytes can assemble and secrete CMs, but cannot exit the gut interstitium. PLAGL2 is involved in CM modification and is necessary for subsequent CM uptake  can lead to lacteal regression without affecting other lymphatic beds, thereby influencing lipid absorption and enhancing excretion of FAs and cholesterol in faces. Vegfc−/− mice exhibited improved glucose metabolism and were protected from HFD-induced obesity  in LECs exhibited shorter lacteals and the survival and migration of IEC were unable to maintain  developed intestinal lymphangiectasia and lacteals uptake lipids defecits and exhibited reduced weight gain  had defective lymphatic vascular integrity, and the accumulation of lymph can lead to adipocyte proliferation and development of spontaneous adult-onset obesity ; ii) lacking clinical safety and efficacy of the dietary TG absorption-related synthetic small biomolecules; iii) unexpected side effects by inhibiting certain key targets, such as severe gastrointestinal reactions , systemic dyslipidemia , and other adverse reactions that interrupt the long-term treatment. Furthermore, differences in sex, living environment, circadian rhythms, and dietary nutrient structure also hunt the therapeutic benefits. Therefore, regulating dietary TG absorption to control HFD-induced obesity is still challenging.Dietary TG absorption involves various regulatory genes and proteins that play a vital role in regulating the related processes. Besides, the length of the carbon chains, the number of unsaturated bonds, and the location of FAs in dietary TG affect absorption efficiency and metabolic phenotypes. The undergoing studies are limited in: i) the expected efficacy is unsatisfied by intervening in certain essential genes or proteins (e.g., CD36, FABPs, DGAT, SAR1A, We note that recently published high-quality studies have indicated some key targets of the intestinal lipid absorption process are promising in therapeutic prospects, such as MGAT2 inhibitors and intestinal-specific MTP inhibitors. TG absorption is an efficient and delicate process. Further research is needed to balance the metabolic benefits and side effects. Combination therapy that regulates multiple targets of intestinal lipid absorption may be an alternative and safe therapeutic approach.


The coronavirus disease 2019 (COVID-19) can cause dermatological manifestations such as urticarial rash, confluent erythematous/maculopapular/morbilliform rash, papulovesicular exanthema, chilblain-like acral pattern, livedo reticularis/racemosa-like patterns, and purpuric “vasculitic” patternsHerein, we describe a case of sarcoidosis with exuberant cutaneous and pulmonary manifestations after recovery from COVID-19 infection. In the literature, only two cases of granulomatous sarcoid-like reaction have been described, with few lesions and no pulmonary involvement of the diseaseA 57-year-old woman with no previous comorbidities was admitted to the dermatology department with erythematous, symmetrical, non-pruritic papules and plaques; some had an annular disposition over six months of evolution A-D. The There may be an intrinsic relationship between viral infections and sarcoidosis development in genetically susceptible hosts


In this review, we aim to discuss the contribution of tumor-associated HA to cancer inflammation, angiogenesis, and tumor-associated immune suppression. We also highlight the recent findings related to the enhanced HA degradation in the tumor microenvironment.Hyaluronan (HA) is known to be a prominent component of the extracellular matrix in tumors, and many solid cancers are characterized by aberrant HA metabolism resulting in increased production in tumor tissue. HA has been implicated in regulating a variety of cellular functions in tumor cells and tumor-associated stromal cells, suggesting that altered HA metabolism can influence tumor growth and malignancy at multiple levels. Importantly, increased HA production in cancer is associated with enhanced HA degradation due to high levels of expression and activity of hyaluronidases . Understanding the complex molecular and cellular mechanisms involved in abnormal HA metabolism and catabolism in solid cancers could have important implications for the design of future cancer therapeutic approaches. It appears that extensive crosstalk between immune cells and HA-enriched stroma contributes to tumor growth and progression in several ways. Specifically, the interaction of tumor-recruited Hyal2-expressing myeloid-derived suppressor cells (MDSCs) of bone marrow origin with HA-producing cancer-associated fibroblasts and epithelial tumor cells results in enhanced HA degradation and accumulation of small pro-inflammatory HA fragments, which further drives cancer-related inflammation. In addition, hyaluronan-enriched stroma supports the transition of tumor-recruited Hyal2 N-acetylglucosamine. Normal physiological HA polymers consist of 2,000-25,000 disaccharides with molecular masses in the range of 106 – 107 kDa and polymer lengths of 2-25µm. HA is synthesized at the cell membrane as an unmodified polysaccharide by one of three hyaluronan synthases (HAS): HAS1, HAS2, or HAS3.Hyaluronan, also called hyaluronic acid (HA), is a member of the glycosaminoglycan family of polysaccharides synthesized at the cell surface. Distributed widely through vertebrate connective, epithelial, and neural tissues, interactions between HA and the extracellular matrix (ECM) regulate cellular processes involved in embryonic development, tissue healing, inflammation, and tumor progression –3. HA isN-acetylglucosamine and glucuronic acid to generate HA fragments , mitogen-activated protein kinases (MAPKs), nuclear factor NF-κB, RAS, phosphatidylinositol kinase (PI3K), tyrosine kinases, and cytoskeletal components , a major component of the extracellular matrix, is anti-oncogenic, which exerts anti-inflammatory and wound-healing activities. Evidence of anti-oncogenic properties of HMW-HA comes from the fact that naked mole rats show unusual resistance to cancer and the maximal lifespan of these mice exceeds 30 years . AnalysiIn contrast, the low-molecular-weight hyaluronan (LMW-HA) is pro-oncogenic. Thus, the HA fragments with low molecular weight produced in a hyaluronidase-dependent manner, inhibited Hippo signaling by competing with HMW-HA for CD44 binding . Tumor g  behavior. One of the first studies demonstrated that partial degradation products of HA produced by the action of testicular hyaluronidase induced an angiogenic response, such as the formation of new blood vessels in the chick chorioallantoic membrane. Further fractionation of the digestion products showed that the activity was restricted to LMW-HA fragments between 4 and 25 disaccharides in length . More reormation –50. Sincormation . Similarormation . AdditioIt appears that enhanced HA degradation is associated with tumor cell spreading. For example, accumulation of LMW-HA in tumor interstitial fluid correlates with lymphatic invasion and lymph node metastasis . Analysivia major pathways including the MAPK  and SRC/FAK pathways that promote expression of an oncogenic transcriptome required for tumor cell survival, migration invasion, proliferation, and resistance to apoptosis  expression/activity . Functiovia cell surface HA receptors. In cancer, the epithelial tumor cells frequently express the membrane-bound Hyal2 and can break down secreted HA into intermediate 20 kDa fragments. Analysis of tumor tissues from cancer patients and experimental animal tumors revealed that tumor cells frequently work together with tumor-recruited myeloid cells including myeloid-derived suppressor cells (MDSCs) and tumor-associated macrophages (TAMs) to break down the extracellular HA into small HA fragments  using various mechanisms primarily from bone marrow and spleen , 80. Upod stroma . Thus, Hd stroma Figure 2d stroma .+ MDSCs have been detected in peripheral blood and tumor tissue in patients with bladder and kidney cancers (+CD33+CD11b+). A similar subset of Hyal2-expressing MDSCs  has been found in tumor-bearing mice  in the tumor microenvironment. In addition, the tumor-infiltrating myeloid cells show significantly higher expression of Hyal1 than tumor cells and have the ability to degrade HA into much smaller fragments with a molecular weight of less than 5 kDa (The increased numbers of Hyal2 cancers , 66. In ing mice . Mobilizan 5 kDa . The smancers +CD3+CD11b+.an 5 kDa , 45.+ MDSCs also co-express the immunosuppressive ligand PD-L1 (+ MDSCs can proliferate and differentiate into immunosuppressive macrophages, forming the large PD-L1+ myeloid cell clusters (+MDSCs into TAMs, the activity of HA-degrading activity of Hyal2 is reduced, while expression of immunosuppressive ligand PD-L1 is markedly up-regulated. It is still unclear how the degradation of extracellular HA promotes differentiation of Hyal2+Gr-1+PD-L1+ MDSCs into more mature immunosuppressive F4/80+PD-L1+ tumor-associated macrophages. It is plausible that under pro-inflammatory conditions in the tumor microenvironment small HA fragments promote the transition of MDSCs into activated immunosuppressive PD-L1+ macrophages. Collectively, these data indicate that tumor-recruited myeloid cells contribute to tumor growth through degradation of tumor-associated HA, promoting cancer-related inflammation, tumor angiogenesis, and tumor-associated immune suppression.Importantly, the tumor-recruited Hyal2nd PD-L1 , 65. In clusters 4. Intervia the PD1/PD-L1 pathway (+ MDSCs, CAFs, and endothelial cells contribute to the highly dynamic development of tumor stroma, enhances degradation of tumor-associated HA, and further promotes the tumor progression through stimulation of cancer-related inflammation, tumor angiogenesis, and tumor-associated immune suppression. Therefore, the normalization of HA metabolism in the tumor microenvironment could potentially provide a strong beneficial step for improving the efficacy of existing approaches to treat cancer, particularly for cancer immunotherapy.Understanding the complex molecular and cellular mechanisms involved in abnormal HA metabolism and catabolism in solid cancers could have important implications for the design of future cancer therapeutic approaches. It appears that extensive crosstalk between tumor-associated myeloid cells and HA-enriched stroma contributes to the tumor growth and progression in several ways. Specifically, the interaction of tumor-recruited Hyal2-expressing myeloid cells with HA-producing stromal cells results in enhanced HA degradation and accumulation of small pro-inflammatory and pro-angiogenic HA fragments . HA-enri pathway , 66. ConThe original contributions presented in the study are included in the article/supplementary materials. Further inquiries can be directed to the corresponding author.WD wrote review materials related to HA metabolism in normal and malignant tissues. PD-G wrote review sections related to the roles of LMW-HA in inflammation and angiogenesis. SK composed the article, wrote review materials related to mechanisms of HA degradation in tumor microenvironment and stroma, wrote abstract, conclusion and prepared Figures. All authors contributed to the article and approved the submitted version.This work was supported by James and Ester King Biomedical Research Program, Florida Health Department, award 8JKO5 and 1923 Fund to SK.SK is a founder of K-Lab Therapeutics.The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Genome-wide association studies (GWAS) have identified a vast number of variants associated with various complex human diseases and traits. However, most of these GWAS variants reside in non-coding regions producing no proteins, making the interpretation of these variants a daunting challenge. Prior evidence indicates that a subset of non-coding variants detected within or near cis-regulatory elements  might play a key role in disease etiology by regulating gene expression. Advanced sequencing- and imaging-based technologies, together with powerful computational methods, enabling comprehensive characterization of regulatory DNA interactions, have substantially improved our understanding of the three-dimensional (3D) genome architecture. Recent literature witnesses plenty of examples where using chromosome conformation capture (3C)-based technologies successfully links non-coding variants to their target genes and prioritizes relevant tissues or cell types. These examples illustrate the critical capability of 3D genome organization in annotating non-coding GWAS variants. This review discusses how 3D genome organization information contributes to elucidating the potential roles of non-coding GWAS variants in disease etiology. Genome-wide association studies (GWAS) have achieved great success during the last two decades, reproducibly identifying hundreds of thousands of genetic variants associated with complex human diseases and traits . HoweverTo better understand the functional roles of non-coding GWAS variants, it is essential to annotate the non-coding regions, which account for ∼97% of the human genome. In recent years, the ENCODE consortium ENCODE  and the The comprehensive annotation of CREs is a substantial step forward in understanding the non-coding GWAS variants. However, it remains challenging to assign non-coding GWAS variants-overlapped CREs to their target genes in disease-relevant tissues and cell types. How CREs regulate the expression of their target genes is still an open question in the genomics field. The difficulties lie in at least four aspects. First of all, the same CRE, such as a super-enhancer, may regulate multiple genes simultaneously. In addition, genes with critical functional roles, such as cell-type-marker genes, may be regulated by multiple CREs simultaneously to allow for some buffer in the presence of disrupted CREs. Along the line, we have recently reported super interactive promoters (SIPs) that interact with a more significant number of CREs than non-SIPs . MoreoveRecently developed genomics and high-resolution imaging technologies  provide This review paper will describe the state-of-the-art experimental technologies, including sequencing-based and imaging-based approaches, to map chromatin spatial organization. In addition, we will summarize advanced computational methods to integrate transcriptome, epigenome, and 3D genome data to achieve a deep understanding of the functional roles of non-coding GWAS variants. We highlight recent breakthroughs in predicting and validating disease causal genes of non-coding GWAS variants and discuss challenges and opportunities for future endeavors.There are three major approaches for examining 3D genome structure: microscopy (imaging)-based techniques, sequencing-based approaches, and integrative methods . Microscin situ hybridization (FISH) and more advanced FISH-based techniques, estimate the relative distance by hybridizing DNA probes of specific genomic regions and then using a microscope for visualization  resolution ; 2021b, 5–106 cells. Single-cell-based sequencing methods, including single-cell Hi-C (scHi-C)  , sci-Hi-(scHi-C) , sc-m3c-(scHi-C)  and Dip-(scHi-C) , all sufTaken together, investigators need to balance the pros and cons of different experimental methods, based on their specific scientific questions. For example, we would recommend imaging-based methods when the primary interest is to understand cell-to-cell variability in regulatory DNA interactions near a specific gene or element of interest. While for another example, when the primary goal is to comprehensively characterize genome-wide enhancer-promoter interactions, sequencing-based methods would be a better choice.in situ genome sequencing (IGS) was designed to jointly conduct sequencing and imaging simultaneously for intact genomes and directly link DNA sequence to 3D spatial proximity  , frequenFTO locus with chromatin interactions identified from 4C-seq  GWAS variant (rs1191551) forming a long-range (∼760 Kb away) interaction with the promoter of FOXG1 revealed by fetal brain Hi-C data (FST and a type 2 diabetes (T2D)-associated SNP rs6450176, which is an intronic variant in ARL15 (BACH2 and rs72928038 (∼30 Kb away), an intronic variant in BACH2 associated with various diseases including multiple sclerosis and type 1 diabetes, detected using promoter capture Hi-C data in naive CD4+ T cells (GATA3 and rs3824662 (∼7 Kb), a GATA3 intronic variant associated with Philadelphia chromosome-like childhood acute lymphoblastic leukemia  disorder. Specifically, the LMNB1 gene becomes highly expressed due to the missing boundary leading to new chromatin interactions between the promoter of the LMNB1 gene and several other enhancers   model, combining chromatin activity and interaction information, assigns rs12740374, a GWAS variant associated with low-density lipoprotein cholesterol (LDL) to the atocytes . We visuatocytes  with a sghlight) . The ABCghlight) .a priori defined genes or region units. The eSCAN method focuses on variants residing in putative enhancer regions, which can increase statistical power by reducing the search space among non-coding regions. Furthermore, with its scan feature, eSCAN tends to identify associated regions that are shorter in size, effectively achieving fine-mapping of causal variants and regions. Integration with chromatin conformation data also makes easier biological interpretation of detected regions. As an aggregation method that tests a set of variants, eSCAN may not be able to narrow down to single variant level. With higher resolution (Kb or finer) chromatin conformation data, eSCAN can potentially pinpoint individual variants. When applied to hematological traits, eSCAN pinpointed multiplied regulatory regions associated with various blood cell indices. These regions were either missed by alternative methods or in much coarser resolution. Among them, a regulator region  was associated with platelet count, a signal missed by standard GWAS. The gene it regulates, the BACH2 gene, is an essential immune cell regulatory factor and plays a critical role in maintaining regulatory T-cell function and B-cell maturation , methods that integrate additional omics data at either bulk tissue or single cell level will further enhance power to prioritize and pinpoint important functional variants, regions and genes, and potentially in tissue- or cell-type-specific manner.Third, studying of chromatin spatial organization can further benefit from advanced machine learning or deep learning methods. Deep learning-based methods have been used for chromatin interaction prediction or Hi-C and alike data enhancement. For example, Akita  adopts aFinally, as a community, we need to generate high quality, high resolution data from complementary technologies in diverse biosamples. First, we need more comprehensive compendia of chromatin conformation data. Such data holds and has been delivering on the promise of helping to fulfill the crucial variant-to-function task. Future efforts should encompass diverse tissues and cell types across developmental stages, multiple disease progression time-points, and under various natural and perturbed conditions, as provided by recent publications , 2020 anInterpretation of GWAS results has received extensive attention in the past two decades, with many alternative approaches proposed and employed to achieve the variant-to-function goal. For example, eQTL and co-localization with GWAS signals , transcr


Acquired von Willebrand disease (aVWD) is frequently observed in patients with the need for extracorporeal membrane oxygenation (ECMO). aVWD can be treated by plasma-derived concentrates containing factor VIII (FVIII) and/or von Willebrand factor (VWF) and recombinant VWF concentrate as well as adjuvant therapies such as tranexamic acid and desmopressin. However, all of these therapeutic options possibly cause thromboembolism. Therefore, the optimal treatment remains uncertain. This report presents a case of a 16-year-old patient suffering from severe acute respiratory distress syndrome due to coronavirus disease 2019 with the need of ECMO support. Our patient developed aVWD under ECMO therapy characterized by loss of high-molecular-weight multimers (HMWM) and severe bleeding symptoms following endoscopic papillotomy due to sclerosing cholangitis. At the same time standard laboratory parameters showed hypercoagulability with increased fibrinogen level and platelet count. The patient was successfully treated with recombinant VWF concentrate  combined with topic tranexamic acid application and cortisone therapy. rVWF concentrate vonicog alfa is characterized by ultra-large multimers and absence of FVIII. Patient could be successfully weaned from ECMO support after 72 days. Multimer analysis 1 week after ECMO decannulation showed an adequate reappearance of HMWM. Hemorrhagic events are frequently observed in critically ill patients with the need for extracorporeal membrane oxygenation (ECMO) therapy.VWF-antigen (VWF:Ag), VWF-collagen-binding-activity (VWF:CB), and VWF-ristocetin-induced-binding (VWF:GP1bR) were determined applying HemosIL AcuStar-Assays  including the calculation of VWF:GP1bR/Ag and VWF:CB/Ag ratio. VWF HMWM were analyzed using the Hydragel 5 von Willebrand Multimer kit and a Hydrasis 2 analyzer .248%) 1 week after reversed phase polymerase chain reaction confirmed SARS-COV-2 infection. Initially, the patient could be stabilized with nasal high-flow therapy and noninvasive ventilation. Sixteen days after ICU admission, the patient had to be intubated, but gas exchange remained persistently insufficient. Due to refractory hypoxemia, therapy with ECMO, cortisone, and prone positioning were initiated and adequate gas exchange could be restored immediately  with severe dyspnea and hypoxia  were hospitalized, 1,423 (0.85%) met criteria for severe disease, and 42 (0.03%) required ECMO.Secondary sclerosing cholangitis (SSC) of the critical ill patient including COVID-19-related cholangiopathy (SC-CIP) is an underestimated and underdiagnosed disease with poor prognosis .This case features the importance of aVWD in ECMO patients, the efficient use of rVWF concentrate in aVWD, and the high-dose methylprednisolone therapy in OP in a pediatric COVID-19 patient. Transfusion of rVWF should therefore be considered as a potential therapeutic strategy for ECMO patients bleeding due to aVWD and high levels of factor VIII at the same time. If OP is suspected a high-dose methylprednisolone therapy should be considered even after initial failure of prednisolone therapy.