Over the last decades, our understanding of adaptive immune responses to solid organ transplantation increased considerably and allowed development of immunosuppressive drugs targeting key alloreactive T cells mechanism. As a result, rates of acute rejection dropped and short-term graft survival improved significantly. However, long-term outcomes are still disappointing. Recently, increasing evidence supports that innate immune responses plays roles in allograft rejection and represents a valuable target to further improve long-term allograft survival. Innate immune cells are activated by molecules with stereotypical motifs produced during injury  or infection . Activated innate immune cells can exert direct pro- and anti-inflammatory effects, while also priming adaptive immune responses. These cells are activated after transplantation by multiple stimuli, including ischemia-reperfusion injury, rejection, and infections. Data from animal models of graft rejection, show that inhibition of innate immunity promotes development of tolerance. Therefore, understanding mechanisms of innate immunity is important to improve graft outcomes. This review discusses effects of currently used immunosuppressive agents on innate immune responses in kidney transplantation. For many years, strategies to prevent allograft rejection have focused purely on preventing adaptive immunity. Recent evidence has increasingly indicated that pure focus on T and B cells is not sufficient to improve long-term renal transplant outcomes –3. InnatInnate immune cells are activated by common mechanisms. Molecules with stereotypical motifs produced during injury  or infection  initiate a variety of inflammatory events, including diapedesis, inflammatory cytokine production, and cell death . These pHerein, we will review relevant literature regarding the impact of the main immunosuppressive agents employed in the maintenance phase of kidney transplantation  on innate immune responses.Calcineurin inhibitors (CNI), such as tacrolimus  and cyclosporine A (CsA), still represent the mainstay of immunosuppression in kidney transplantation. Their dominant mechanism of action is the inhibition of Nuclear Factor of Activated T-cells (NFAT) phosphorylation, with consequent reduction of IL-2-mediated T lymphocyte activation and proliferation , 11. CNIEvidence shows that CsA blocks NFAT binding to the inducible NO synthase (iNOS) promoter, causing a reduction of iNOS expression and nitrite production in macrophages , 15. CsACsA reduces the secretion of pro-inflammatory cytokines tumor necrosis factor (TNF)-α and IL-12 induced by LPS in human DCs –20 and mThis effect could be enhanced in patients treated with anti-thymocyte globulins. As reported by Naujokat et al. , DCs areCNI have also an inhibitory effect on Toll-like receptors (TLRs) dependent activation of monocytes/macrophages. In monocytes/macrophages from liver transplant recipients, therapeutic concentrations of CsA impaired IL-6 production in response to TLR2 and TLR7/8 activation, and TNF-α synthesis due to TLR7/8 stimulation, more than TAC . In renaMoreover, although there are not conclusive findings regarding the effects of maintenance immunosuppressive drugs on innate immunity and their impact on ischemia reperfusion, the study of Yang et al.  suggesteCNI may also influence immune cells by affecting their mitochondrial function . In macr−/− mice, lacking B and T cells, CNI treatment induced a rapid development of Candida albicans infections, indicating that CsA impairs specific anti-fungal functions in innate immune cells  , causingne cells . More sprocesses .Currently, overall mortality due to fungal infections in transplant patients varies between 25 and 80%, with Candida and Cryptococcus species being the most commonly identified yeasts .Ex vivo studies revealed that CsA damages human neutrophil clearance of Aspergillus fumigatus (another important cause of post-transplant opportunistic infections)  , and thaCNI do also affect NK cells in kidney transplant recipients . Zhang eex vivo treatment of NK cells from healthy controls with CNI inhibits their degranulation and IFN-γ production. Similar functional impairment was observed in NK cells from CNI-treated patients. This could have dramatic effects on the NK cells capacity of killing transformed or virus-infected cells and producing pro-inflammatory cytokines and could, at least in part, explain the increased risk of opportunistic infections and tumors of CNI-treated patients (CsA inhibits the NK cells proliferation in a dose-dependent manner . Morteaupatients .de novo guanosine nucleotide synthesis by selectively inhibiting the inosine monophosphate dehydrogenase (IMPDH), mainly expressed by T- and B- cells  and its active metabolite mycophenolic acid (MPA), are the most widely used drugs in transplantation , 40. MMFB- cells , 42.When exposed to MMF/MPA, monocytes show lower levels of pro-inflammatory cytokine IL-1β and altered polarization, with enhanced expression of surface markers (like CD163 and CD200R), generally associated with an anti-inflammatory function (M2 phenotype) . AdditioIn a mouse model of renal IRI, MMF down-regulated TLR4 expression on monocytes surface, along with plasma level of several cytokines . This resulted in milder kidney damage, as defined by creatinine levels and histological findings at 48 h after IRI .MMF also reduces the LPS-induced expression of MHC-II on monocyte surface, suggesting a reduced activity as antigen presenting cells . In the MPA and MMF have similar effects on hMDDCs activation and maturation, but MMF reduces, instead of increasing, IL-10 synthesis. This may support the concept that MPA has stronger protolerogenic effects on monocytes compared to MMF . It is lMMF/MPA have also modulating effects on NK cell activity. Similarly to mTOR inhibitors, they significantly reduce the proliferation of these cells and inhibit the expression of CD56, associated with a highly reactive phenotype , 51. AccTaking together, these data suggest that MMF/MPA impair differentiation, maturation and function of various innate immunity cells, which may represent an additional mechanism of their immunosuppressive effects. Whether similar mechanisms are shared with azathioprine, an antiproliferative agents with similar antirejection effects , is unclGlucocorticoids (GCs) are anti-inflammatory drugs employed in both induction and maintenance phase of immunosuppression after kidney transplantation. They inhibit the inflammatory response and leukocyte migration into inflamed tissues. They also accelerate resolution of inflammation by inhibiting vascular permeability and leukocyte distribution/trafficking, and by modulating death/survival and cellular differentiation programs , 54.Until recently, it has been thought that the anti-inflammatory effects of GCs were linked to their ability to inhibit regulator of genes encoding pro-inflammatory cytokines  through a mechanism called “transrepression” . HoweverIn vitro, methylprednisolone-treated monocytes show increased expression of anti-inflammatory cytokines, like IL-10, with concomitant down-regulation of TNF-α, IL-1β, and IL-12  and CD14++CD16+ (intermediate) monocytes, while the CD14+CD16++  population is declined compared to patients receiving CNI, MMF/MPA or mTOR inhibitor (in vitro expression of TLR4 and the response to endotoxin in monocytes through the mediation of micro-RNA (MiR) 511-5p, a keystone in the anti-inflammatory effect of GCs  is part of 2 different complexes (mTORC1 and mTORC2) with diverse signaling networks. mTORC1 promotes anabolic cellular metabolism stimulating synthesis of proteins, lipids, and nucleotides and, at the same time, inhibits catabolic processes, such as lysosome biogenesis and autophagy. mTORC2 controls cell survival, cytoskeleton organization, lipogenesis, and gluconeogenesis . In orgaThe mTOR network allows innate immune cell maturation and costimulatory molecule expression during inflammation . As mighConversely, mTOR inhibitors indirectly inhibit regulation of autophagy and promote this degradation with immunoregulatory capabilities. Importantly autophagy is a well-known contributor to both MHCII presentation and MHCI cross-presentation of exogenous peptides , 97. IncIn immature DCs, mTOR inhibitors induce apoptosis by blocking the granulocyte-macrophage colony-stimulating factor (GM-CSF) signaling. Disruption in GM-CSF/PI3K/mTOR pathway produces a pro-apoptotic state, unbalancing anti- and pro-apoptotic mediators by reduction of the mitochondrial membrane potential . In matubrightCD16+/− NK cell subset (associated with enhanced cytokine production) without affecting the amount of CD56dimCD16+ cells subset (with more cytotoxic capacity). With regards to the cytotoxic subset, mTOR inhibitors prevented NK cell expression of NKG2A and NCR (mTOR inhibitors inhibit NK cell inflammatory capabilities by inhibiting their cytokine expressing and cytotoxic function. In particular, rapamycin impaired growth of the CD56 and NCR . Absence and NCR .An essential innate immune cell role involves production of cytokines. The mTOR inhibitors have pleiotropic effects that depend on the cells and circumstances studied. In LPS-stimulated human monocytes, mTOR inhibitors reduce several pro-inflammatory chemokines synthesis such as MCP-1, RANTES, IL-8, and MIP-1 . Fine-nemTOR inhibitor modulation of innate immune cells may contribute to a pro-tolerogenic state in the early phases of transplantation. Sordi et al.  showed tThere are controversial data about the impact of the m-TOR inhibitors on the regulation of I/R injury-related innate immune system in kidney transplantation. Several authors have suggested that these drugs may impair recovery of kidney function –114 becaIn the past, great strides in allograft survival prolongation were attributed to successful suppression of adaptive immune responses . A greatGZ, JL, and PC searched the literature and wrote the manuscript. LS contributed to the literature search and literature analysis. GG and PC revised the manuscript. All authors read and approved the final manuscript.The 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.


Carbamylation is a post-translational modification that mostly affects proteins with low turnover, such as dermal proteins. Carbamylated proteins accumulate in skin in an age-dependent manner, contributing to tissue alterations. As dermis is affected by systemic sclerosis (SSc) and anti-carbamylated protein antibodies (anti-CarP Ab) are found in SSc patients, we sought to evaluate the specificity of anti-CarP Ab and their relationship with clinical parameters reflecting skin involvement in SSc. This study investigated serum samples and clinical data from 124 patients with SSc. Anti-CarP Ab were affinity purified from pooled SSc sera, and their specificity was assessed by western blotting and ELISA with carbamylated proteins from two species . Anti-CarP Ab were measured in SSc serum samples and in 41 healthy aged-matched individuals. Affinity-purified anti-CarP Ab recognized carbamylated epitopes irrespective of the protein type or species origin. Anti-CarP Ab levels inversely correlated with the modified Rodnan skin score (mRss) , independently of patients’ age. Receiver operating characteristics (ROC) analysis identified anti-CarP Ab cut-offs that best discriminated dichotomized clinical variables related to skin involvement: the only clinical variables that were significantly different between groups were mRss (p = 0.001) and scleredema (p<0.001). Low anti-CarP Ab levels were associated with worse skin involvement. Future prospective studies are needed to assess their usefulness in the clinical setting. Systemic sclerosis (SSc) is a rare, multisystem, connective tissue disease of unknown etiology and pathogenesis, characterized by three interconnected pathogenic events, namely vascular abnormalities, abnormal extracellular matrix (ECM) deposition, and autoimmunity ,2. ThougPatients with SSc present auto-antibodies (Ab) with a wide range of specificity. Circulating antinuclear Ab directed to self-antigens such as DNA topoisomerase I, RNA polymerase III, the Th/To autoantigen, and heterologous centromeric proteins (CENPs) have been detected in over 95% of patients , and havCitrullination is the enzymatic conversion of peptidyl-arginine into peptidyl-citrulline. CarbamyPrevious studies identified various antigens that are targeted by Ab against carbamylated proteins (anti-CarP Ab), namely albumin , hemogloConsidering that dermis is one of the main targets of SSc and that dermal proteins undergo carbamylation, here we assessed whether the neo-carbamylated epitopes recognized by anti-CarP Ab in SSc are conserved among different proteins from different species and whether the levels of these Ab are associated with clinical manifestations related to skin involvement in SSc.+ (n = 56) and anti-Scl70 Ab+ (n = 55).124 SSc patients who fulfilled both the 1980 ACR  and the This study was approved by the Ethics Committees of the Universities of Naples, Bari and Foggia, and written informed consent was obtained from each participant.Bovine serum albumin (BSA) and human fibrinogen (FIB) were purchased from Sigma-Aldrich. Human albumin (HA) was purchased from Kedrion Biopharma . Electrophoresis reagents were purchased from Bio-Rad Laboratories. Horseradish-peroxidase (HRP)-conjugated xeno-Ab to human IgG (Fc portion) was purchased from Jackson Immunoresearch Laboratories .Serum samples from the 124 SSc patients and from 41 age-matched healthy blood donors (HBD) were stored at -80°C until use.Carbamylated BSA (CarBSA), human albumin (CarHA), and human fibrinogen (CarFIB) were generated by incubating 2 mg protein in 1 ml 8 M urea, 100 mM Tris-HCl (pH 8.5), for 15 h at 61°C. After extensive dialysis against phosphate-buffered saline (PBS), carbamylated protein concentration was assessed using the BCA assay (Thermo Fisher Scientific). Success of the carbamylation procedure was verified by documenting a reduced electrophoretic mobility  by SDS-Po-phenylenediamine. Specific binding was determined by subtracting the background binding in BSA-coated wells from the binding in experimental wells. The levels of anti-CarP Ab in sera were expressed as a percentage of binding relative to that of the positive control. Samples were tested in duplicate, and the experiment was performed at least 3 times. Ten serum samples with highest titer were pooled and used for affinity purification of anti-CarP Ab. The specificity of anti-CarP Ab was assessed by ELISA in binding and inhibition assays as previously described  of anti-CarP Ab between SSc (28.17±25.43) and HBD (34.24±24.47) groups  and with mRss . Because lower anti-CarP Ab were observed in the oldest age group (≥67 years), we repeated Spearman's analysis after group V exclusion. Even with this exclusion, the association between anti-CarP Ab and mRss remained statistically significant (p = 0.008), suggesting that the inverse association between anti-CarP Ab and mRss is not influenced by age. For this reason, we decided to use the whole population for further investigations.Spearman's correlation analysis was performed to define the relationship between serum anti-CarP Ab levels and age or mRss in SSc patients. As reported in + (25.72±18.93)and patients with anti-Scl70 Ab .We also assessed a possible relationship between anti-CarP Ab and SSc-specific auto-Ab, namely ACA and anti-Scl70 Ab. This analysis revealed no significant differences in the mean levels [± standard deviation] of anti-CarP Ab between patients who were ACAFig 4A) and to the presence or absence of scleredema . The best anti-CarP Ab cut-off discriminating patients with mRss>0 (versus those with mRss = 0) was ≤23.5% of the binding , while the best cut-off discriminating patients with scleredema from those who did not have it was ≤25.6% . These results indicate that anti-CarP Ab are inversely associated with worse skin involvement in SSc patients.ROC curves were generated for anti-CarP Ab to evaluate their ability to discriminate dichotomized clinical variables describing skin involvement in SSc. Acceptable ROC curves (p<0.05) were obtained only with the dichotomization of SSc patients according to mRss (vs>0) and scleredema (absence vs presence) as outcome variables, and anti-CarP Ab levels (log-transformed) along with sex, age at diagnosis, age at observation, disease duration (log-transformed) and clinical type (limited or diffuse) as predictor variables. As shown in Table 4, low levels of anti-CarP Ab were associated with mRss>0 and the presence of scleredema, indicating that these Ab inversely associated to the severity of skin involvement, independently of disease duration, both in the whole cohort and in the age group <67. Disease type (limited vs diffuse), sex, age at diagnosis and age at observation remained excluded from the model. When “age at observation” was used as the outcome variable, anti-CarP Ab levels were not retained in the model, supporting the lack of association between these two variables.Finally, to further analyze the relationships between anti-CarP Ab and skin involvement and its interdependency with age, multivariate forward linear regression was performed by including mRss (score 0 In the present study, we characterized the specificity of Ab directed against carbamylated proteins in SSc. We demonstrated that the recognition of carbamylated proteins by anti-CarP Ab occurs by means of the carbamylation-dependent epitope, irrespective of the species origin or type of protein.in vitro carbamylation of fibrinogen modified all its three distinct bands, but the anti-CarP Ab reacted strongly only with the α chain. Similar western blotting results have previously been reported by Shi et al., who used a different procedure to carbamylate fibrinogen and who tested anti-CarP Ab from RA sera [The  RA sera . The lacIn this study, we found no differences in serum levels of anti-CarP Ab between SSc patients and normal healthy subjects. This finding is not unique, since similar results were obtained by Pecani et al. , who meaWe found a progressive, age-dependent decrease in anti-CarP Ab serum levels in our patient cohort, in line with the reported age-dependent accumulation of carbamylated dermal collagen . The decThe mechanisms underlying the associations between low anti-CarP Ab levels and the severity of skin involvement, reflected by mRss>0 and scleredema, can only be speculated at the present. One possibility is that high amounts of carbamylated proteins in skin, the target of carbamylation, may actFinally, all anti-CarP Ab ELISAs performed so far , 25, lik


Human cytomegalovirus (HCMV) reactivation is a major infectious cause of morbidity and mortality after allogeneic hematopoietic stem cell transplantation (HSCT). HCMV is a ubiquitous beta-herpesvirus which asymptomatically infects immunocompetent individuals but establishes lifelong latency, with the potential to reactivate to a life-threatening productive infection when the host immune system is suppressed or compromised. Opportunistic HCMV reactivation is the most common viral complication following engraftment after HSCT and is associated with a marked increase in non-relapse mortality, which appears to be linked to complex effects on post-transplant immune recovery. This minireview explores the cellular sites of HCMV latency and reactivation in HSCT recipients and provides an overview of the risk factors for HCMV reactivation post-HSCT. The impact of HCMV in shaping post-transplant immune reconstitution and its relationship with patient outcomes such as relapse and graft-versus-host disease will be discussed. Finally, we survey current and emerging strategies to prevent and control HCMV reactivation in HSCT recipients, with recent developments including adoptive T cell therapies to accelerate HCMV-specific T cell reconstitution and new anti-HCMV drug therapy for HCMV reactivation after HSCT. Human cytomegalovirus (HCMV) is a beta-herpesvirus carried by a majority of the global population . The serHCMV reactivation is the major viral infectious complication after allogeneic HSCT and is associated with an increased risk of non-relapse mortality , which iThe cellular sites and mechanisms associated with HCMV latency and reactivation are incompletely understood, in part due to the high human-specificity of HCMV which precludes the use of animal models to directly study HCMV infection. Clinical evidence indicates that the risk of reactivation post-HSCT is strongly connected to the pre-transplant HCMV serostatus of the donor and recipient  and the Allogeneic HSCT is the only curative option for many hematological malignancies and diseases . GvHD, ode novo T-cell reconstitution via the thymus from donated pluripotent hematopoietic stem cells to generate a primary T-cell response to HCMV reactivation. By contrast, HCMV-specific memory T-cells contained in D+ grafts can undergo more rapid antigen-driven expansion within the recipient in the early post-transplant period and contribute to early control of reactivation (Reactivation develops in over 60% of HCMV seropositive recipients (R+), and in approximately 10% of seronegative recipients (R-) transplanted from seropositive donors (D+) . In R+/Dtivation . For a stivation , 2014. NAdvances in transplant and antiviral treatment practices over the last 25 years have reduced the incidence of HCMV disease to ∼10% in the first year post-transplant . However+ cells and CD33+ myeloid progenitor cells (+ monocytes and myeloid dendritic cells (mDCs)  . A receny donors . It may  latency .Latently infected cells contain HCMV DNA  but do n+ cell subset (Latently infected cells are present at very low frequencies (0.004–0.01% of mononuclear cells) in GCSF-mobilized peripheral blood or bone-marrow from healthy seropositive donors , but undl subset  also reml subset  and recel subset . The speIn addition to recipient and donor HCMV serostatus, independent risk factors for reactivation include increasing recipient age , use of + and CD8+ T-cells expand with reactivation and are likely both required for control and/or protection  γδ T-cell proliferations suggestive of adaptive responses (+ T-cell repertoire diversity in the elderly (Reactivation stimulates the recovery of HCMV-specific T-cells after HSCT (see section “Risk Factors for HCMV Reactivation After HSCT”) , but is ansplant , which i T-cells , leadingtivation . The sel T-cells , and cloesponses , are alsesponses . Lugtharesponses , althoug elderly . The pro elderly , but the+ NK cells (dimCD57+NKG2A-CD158b+) NKG2C+ NK cells persist after viral clearance (+CD57+ NK cells are also observed in R+/D+ patients with subclinical HCMV infection (+ NK cells of HCMV UL40 peptides presented in the context of HLA-E was recently identified to be the mechanism that drives the expansion and differentiation of NKG2C+ NK cells during HCMV infection (HCMV reactivation drives a rapid expansion of IFN-γ-producing NKG2CNK cells , which lNK cells . Expandelearance  and memonfection . Specifinfection .dimCD57+NKG2C+ NK cell and Vδ2neg γδ T-cell subsets which expand with reactivation (+ T-cells (An association between early HCMV reactivation and reduced myeloid leukemia relapse has been reported . This putivation , or via  T-cells . However T-cells . Furtherex vivo (Graft-versus-host disease and its steroid therapy increase the risk of reactivation after HSCT . The allex vivo . Conversex vivo  and the ex vivo . An increx vivo . Furtherex vivo  might plStandard antiviral drugs for reactivation after HSCT are ganciclovir, valganciclovir and foscarnet. Prophylactic use is reserved for high-risk patients due to their significant toxic side effects. Ganciclovir causes neutropenia which increases the risk of bacterial and fungal superinfections , while fProphylactic administration of Letermovir, a new anti-HCMV agent which inhibits the viral terminase complex, recently demonstrated the capacity to reduce the risk of HCMV disease and all-cause mortality at 24 weeks post-transplant, in a Phase 3 trial . There aPre-emptive treatment based on viral DNAemia surveillance  minimizeex vivo or isolated directly with HCMV-specific tetramers and infused in the post-transplant period can accelerate HCMV-specific immune recovery and contribute to long-term control of reactivation and protection from HCMV disease (Pooled HCMV-specific or polyclonal intravenous immunoglobulin is not effective at preventing reactivation or reducing mortality when used in the treatment of HCMV pneumonia post-HSCT , althoug disease . The pos disease . FurtherHCMV is a highly prevalent, opportunistic pathogen that continues to cause substantial morbidity and mortality after HSCT. Improved knowledge of the cellular sites of HCMV latency and the conditions which enable its reactivation to clinically significant infection will be needed to better predict, prevent and control reactivation post-HSCT. Future strategies might involve the selective depletion of latently infected cells from the graft , 2017, pLS and BW generated the initial draft of the manuscript. All authors contributed to the subsequent writing and review of the manuscript.The 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.


Trans-synaptic neurotransmission of both electrical and neurochemical information in the central nervous system (CNS) is achieved through a highly interactive network of neuron-specific synaptic proteins that include pre-synaptic and post-synaptic elements. These elements include a family of several well-characterized integral- and trans-membrane synaptic core proteins necessary for the efficient operation of this complex signaling network, and include the pre-synaptic proteins: (i) neurexin-1 (NRXN-1); (ii) the synaptosomal-associated phosphoprotein-25 (SNAP-25); (iii) the phosphoprotein synapsin-2 (SYN-2); and the post-synaptic elements: (iv) neuroligin (NLGN), a critical cell adhesion protein; and (v) the SH3-ankyrin repeat domain, proline-rich cytoskeletal scaffolding protein SHANK3. All five of these pre- and post-synaptic proteins have been found to be significantly down-regulated in primary human neuronal-glial (HNG) cell co-cultures after exposure to Bacteroides fragilis lipopolysaccharide (BF-LPS). Interestingly, LPS has also been reported to be abundant in Alzheimer’s disease (AD) affected brain cells where there are significant deficits in this same family of synaptic components. This “Perspectives” paper will review current research progress and discuss the latest findings in this research area. Overall these experimental results provide evidence (i) that gastrointestinal (GI) tract-derived Gram-negative bacterial exudates such as BF-LPS express their neurotoxicity in the CNS in part through the directed down-regulation of neuron-specific neurofilaments and synaptic signaling proteins; and (ii) that this may explain the significant alterations in immune-responses and cognitive deficits observed after bacterial-derived LPS exposure to the human CNS. As the basic structural and functional components for inter-neuronal communication, synapses with sufficient, and consistent protein quality and quantity are essential for neural connectivity and functionality in the central nervous system (CNS) to maintain the continuous flow of functional neural information . Therefotrans-synaptic communication resulting in pathogenic neurotransmission that is accompanied by deficits in behavior, cognition, and memory formation.Over the last several years multiple pre-synaptic and post-synaptic proteins including neurexin (NRXN), the synaptosomal-associated phosphoproteins SNAP-25 and synapsin-2 (SYN-2), the type 1 cell adhesion protein neuroligin (NLGN) and the proline-rich SH3-ankyrin repeat-containing cytoskeletal scaffolding protein SHANK3 have been identified and characterized: (i) as being critical to synaptic integrity, acting as key players in the modulation of synaptic neurotransmission, inter-neuronal signaling and synaptic plasticity; and (ii) as being down-regulated in AD and other progressive and lethal inflammatory neurodegenerative disorders of the human CNS . IncreasHomo sapiens contains a complex microbiome consisting primarily of bacteria, with archaea, fungi, microbial eukaryotes, protozoa, viruses, and other microorganisms making up the balance , with Actinobacteria (∼3%), Proteobacteria (∼1%), and Verrucomicrobia (∼0.1%) making up the remaining divisions. These five major bacterial phyla represent the “microbial-core” of the human GI-tract microbiome , as a normal commensal microbe of the human GI-tract, are thought to be usually advantageous to human health due to their abilities to biosynthesize and/or metabolize dietary fiber, complex sugars and polysaccharides, volatile fatty acids, and other nutrients, to function in the development, maintenance, and homeostasis of the host immune and digestive systems  they can cause substantial inflammatory pathology both systemically and within vulnerable CNS compartments -tract microbiome to human neurological health and disease . The GI- balance . Togethe balance . Microbi balance . Interescrobiome . The vasd-to-one . Interes systems . Howeverartments .Bacteroides fragilis (BF-LPS) is one of the most neurotoxic and pro-inflammatory lipoglycans known  cells and preparation of LPS-enriched extracts from different Gram negative bacterial sources has been previously described in detail by our laboratory ,b. The c2+-dependent neurexin/neuroligin (NRXN/NLGN) complexes at CNS synapses; these are required for efficient neurotransmission and are involved in the formation of stable synaptic contacts; NRXN-1 was detected using a NRXN1 ELISA kit; Catalog Number ABIN2870172 (LifeSpan BioSciences); with a detection range of 0.312–20 ng/ml). The synaptosome associated protein 25  is a 25 kDa synaptic vesicle membrane docking and fusion protein whose actions are mediated by SNAREs  located on the synaptic vesicle membrane (v-SNAREs) and/or the target membrane (t-SNAREs); SNAP25 is a pre-synaptic plasma membrane protein involved in the regulation of neurotransmitter release; SNAP25 proteins were detected using a SNAP25 ELISA kit; Catalog Number LS-F17747-1 (LifeSpan BioSciences); with a detection range of 0.78–50 ng/ml. Synapsin-2  is a ∼54 kDa member of the synapsin gene family, encoding a neuronal phosphoprotein that associates with the cytoplasmic surface of synaptic vesicles; SYN-2 is implicated in synaptogenesis and the impairment of neurotransmitter release in multiple neuropsychiatric diseases  is a highly conserved 42 kDa polypeptide that polymerizes to produce microfilaments that form cross-linked networks in the cellular cytoplasm; interestingly β-actin is the most abundant 6 nm diameter microfilament in the synapse; β-actin proteins and filaments were detected using a ACTB/β-actin ELISA Kit; catalog number LS-F10737  with a detection range of 0.312–20 ng/ml. The neurofilament-light (NF-L) chain protein  is a 68 kDa polypeptide that forms a 10 nm diameter type IV intermediate filament specific to the neuronal cytoplasm; together with microfilaments and microtubules NF-L forms the neuronal cytoskeleton and regulates axonal caliber and axonal conduction velocity; NF-L has long been known to exhibit significantly reduced expression in AD brain ; NF-L fidiseases ; SYN-2 welopment ; NLGN waasticity ; levels The neuron-specific NF-L intermediate filament protein, the pre-synaptic proteins NRXN-1, SNAP-25, SYN-2, and the post-synaptic proteins NLGN and SHANK3 constitute a critical mass of axonal filament support elements and synaptic signaling proteins whose deficits might be expected to lead to the inability of neurons and synapses to carry out both electrical- and neurochemical-based neurotransmission in the CNS, and hence contribute to deficits in cognition and memory formation in AD and in neurodegenerative disease models . Why the(i)specific LPS-induced destabilizing effects on cell-adhesion proteins and vascular endothelial cells ;(ii)LPS-mediated generation of ROS and the induction of significant amounts of oxidative stress which directly impacts the abundance and integrity of synapses, synaptogenesis and cognition in murine and related rodent models of neurodegeneration ;(iii)LPS-mediated disruption and opening of the BBB in transgenic AD mouse models ;(iv)LPS- and ROS-induced NF-kB (p50/p65) signaling, known to be a strong inducer of pro-inflammatory microRNAs (miRNAs) which target the 3′-untranslated regions (3’-UTR) of highly selective, AD-relevant mRNAs causing them to be down-regulated ; interes(v)32P]-uridine triphosphate (108 dpm/ml) into newly synthesized total RNA indicates that LPS at nanomolar concentrations strongly inhibits the transcriptional output of neuronal nuclei; this may contribute in part to the generalized down-regulation of gene expression for transcription factors and synaptic and neurotrophic markers as is widely observed in sporadic AD LPS, via synaptogenic targeting, has long been known to contribute to the impairment, alteration or shutting down of essential brain cell cognitive functions such as object recognition and spatial memory formation ;(vii)the progressive cognitive and functional impairment in AD and related forms of progressive inflammatory neurodegeneration is a reflection of progressive neuronal atrophy and synaptic loss; recent preclinical data suggests that LPS-activated microglia may contribute to the elimination of viable neurons and synapses by promoting a neurotoxic astrocytic phenotype known as A1, which can facilitate synaptic disruption and neuro-inflammation in response to peripherally applied LPS ;(viii)a well-documented LPS-mediated neuro-inflammation, apoptosis, BBB disruption, synaptic dysfunction, and cognitive impairment, in multiple murine models of inflammatory neurodegeneration ;(ix)LPS-directed synaptic dysfunction that may be related to the degradation of essential neuronal functions and/or part of a microbial immune-accommodation or evasion strategy directed toward immune tolerization and/or the triggering of an autoimmune response ;(x)prenatal exposure to LPS that results in cognitive deficits in aging of offspring and LPS-induced cognitive impairment, neuro-inflammation, apoptosis, physiological barrier alteration, and synaptic dysfunction in multiple experimental animal models ;(xi)other yet unrecognized LPS-triggered neurological disruptions which significantly disturb homeostatic synaptic function that may be synergistic with other pathogenic factors involved in amyloidogenesis, neuronal atrophy and/or synaptic failure ; or  are targeted by LPS and drive inflammatory neurodegenerative disease; and (ii) create specific alterations in neuronal synaptic transmission, and by doing so contribute to cognitive deficiencies characteristic of AD and the neurodegenerative disease process.In summary, these combined results provide the first evidence that strongly pro-inflammatory, GI-tract microbiome-derived LPS induces both pre-synaptic and post-synaptic changes via the down-regulation of critical and specific synaptic components in primary HNG cell co-cultures, many of which are also observed in AD brain. The findings further support our idea that cytoskeletal-associated and synaptic molecules known to play essential roles in All datasets generated for this study are included in the manuscript and/or the supplementary files.All acquisition, handling, experimental and analytical procedures involving human brain cell cultures (HNG cells) were carried out in an ethical manner in accordance with the ethics review board policies at brain and tissue donor institutions and at the Louisiana State University (LSU) Health Sciences Center. The ethical use of post-mortem human brain cells and their analyses were also carried out in strict accordance with the Institutional Biosafety Committee and the Institutional Review Board Committee (IBC/IRBC) ethical guidelines IBC#18059 and IRBC#6774 at the LSU Health Sciences Center, New Orleans, LA, United States. The work in this research study was reviewed and approved by the IBC/IRB at the LSU Health Sciences Center, New Orleans, LA, United States.WL compiled the research findings and wrote the final version of the manuscript. All authors performed the experiments, organized and compiled the data, and performed the statistical and bioinformatics analysis, and the literature searches.The 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.


Epithelial mesenchymal transition (EMT) is a key process in the progression of malignant cancer. Therefore, blocking the EMT can be a critical fast track for the development of anticancer drugs. In this paper, we update recent research output of EMT and we explore suppression of EMT by natural anti-inflammatory compounds and pro-resolving lipids. The epithelial–mesenchymal transition (EMT) is defined as a phenomenon that epithelial cells transform into mesenchymal cells . EMT plaInflammation is one of ten in cancer hallmark . It is aIn this review, I have dealt with ingredients derived from natural products that were not covered in the 2018 review on EMT . I have EMT shows the reduced expression of epithelial markers including E-cadherin and keratins and the increased expression of mesenchymal marker proteins such as vimentin and N-cadherin via actions of transcription factors including as SNAIL1 and ZEB1  6]. The. The6]. EMT occurs by a various mediators from tumor microenvironments via receptor through signal transduction. EMT-related transcription factors blocks the expression of epithelial cell-marker genes and evoke mesenchymal-marker genes. E-cadherin, keratin, ZO-1, miR-34, and miR-200 belong to the epithelial markers, and N-cadherin, vimentin, fibronectin, SNAIL, ZEB1, TWIST, Brachyury, Foxq1, Runx2, GATA, and SOX belong to the mesenchymal markers. P-cadherin is the marker of partial EMT. The dot triangle indicates the relative ratio of p-cadherin expression. Modified from Lee’s report .A typical signaling pathway of EMT is the transforming growth factor-β1 (TGF-β1) pathway. TGF-β1 induces EMT via SMAD-dependent or non-SMAD signaling pathway . Growth Novel players are newly recognized as regulatory transcription factor in the EMT. Brachyury, the T-box transcription factor, is a novel transcription factor implicated in the EMT of cancer cells . BrachyuEMT could be not defined as a dichotomous transition from epithelial status to mesenchymal one of cells ,25.Partial EMT with both epithelial and mesenchymal cell markers was proposed  25]. Ce. Ce25]. A cancer stem cell (CSC) is a cancer cell having the ability of self-renewal and differentiation. It divides to progenitor cancer cells. It is a culprit of cancer recurrence and metastasis . SeveralIn the course of EMT, epithelial cancer cells can lose contacts between cells and apicobasal polarity but gain enhanced migration and invasion ,6. EMT hChemoresistance to anticancer chemotherapeutics implies that cancer cells can survive despite the administration of an anticancer drug in a dose that can usually kill cancer cells. A significant correlation has been found between EMT-related gene expression and chemo-resistance to anticancer therapy ,30,31. A+ Foxp3+ cells are expanded . H. HOncorhToxoplasma gondii, which in turn reduces IL-12 production by dendritic cells, thus dampening Th1-type cell-mediated immune responses (Infectious organisms can produce pro-resolving lipids to control host inflammation. Thus, supraphysiological levels of LxA4 are generated during infection by esponses  310]..ToxoplasCandida albicans can modulate host defense by biosynthesizing the pro-resolving lipid RvE1 . T. TCandidand RvE2  312]. T. TCandidSeveral natural compounds have anti-inflammatory activities and/or anti-EMT activities. However, few reports have explained their anti-EMT activity by anti-inflammatory or pro-resolving mechanisms. Therefore, examining anti-EMT activities of natural compounds based on their anti-inflammatory or pro-resolving activities and assuring their anti-EMT activities in vivo might be important in the future. Especially, it might be a reasonable way to study the anti-EMT activity of natural products through interaction with the tumor microenvironment .The flux of inflammatory or pro-resolving lipids from arachidonic acid related pathway  could chExpansion of studies about other pro-resolving lipids and nature-derived pro-resolving compounds to inhibit EMT are needed. To do this, various types of pro-resolving lipids and new compounds from natural sources should be procured and diffused to researcher without barrier. Thus, collaboration of natural chemistry researchers with other fields is also required.


Immune cells are one of the most versatile cell types, as they can tailor their metabolic activity according to their required function. In response to diverse environmental cues, immune cells undergo metabolic reprogramming to support their differentiation, proliferation and pro-inflammatory effector functions. To meet a dramatic surge in energetic demand, immune cells rewire their metabolism to utilize aerobic glycolysis. This preferential use of glycolysis even under aerobic conditions is well established in tumor cells, and is known as the “Warburg effect.” Tumor cells avidly use glucose for aerobic glycolysis, thereby creating a nutrient-starved microenvironment, outcompeting T cells for glucose, and directly inhibiting T-cell anti-tumoral effector function. Given that both immune and tumor cells use similar modes of metabolism in the tumor stroma, it is imperative to identify a therapeutic window in which immune-cell and tumor-cell glycolysis can be specifically targeted. In this review, we focus on the Warburg metabolism as well as other metabolic pathways of myeloid cells, which comprise a notable niche in the tumor environment and promote the growth and metastasis of malignant tumors. We examine how differential immune-cell activation triggers metabolic fate, and detail how this forbidding microenvironment succeeds in shutting down the vigorous anti-tumoral response. Finally, we highlight emerging therapeutic concepts that aim to target immune-cell metabolism. Improving our understanding of immunometabolism and immune-cell commitment to specific metabolic fates will help identify alternative therapeutic approaches to battle this intractable disease. Immunometabolism describes the interplay between immunologic and metabolic processes, and can be interpreted in two possible ways: (1) the role of metabolic changes in immune cells in influencing cellular functions and broader immunity , and (2)Immune cells respond to environmental cues  to assume a wide variety of functional states. Immune cells are a vital component of the body’s defense against disease and are important in maintaining tissue homeostasis. Upon encountering a stimulus, immune cells may migrate, proliferate, secrete cytokines and undergo apoptosis. Different immune-cell subsets utilize different metabolic pathways to generate the required cellular energy and biosynthetic macromolecules to fulfill their effector functions when mounting a host immune response.Thus far, the metabolic pathways utilized by T cells have been the most well studied and recently reviewed . As suchMetabolic reprogramming is a hallmark of cancer . For tumCells can also use glutamine or fatty acids as nutrients to fuel the TCA cycle . For exaOXPHOS is a highly efficient pathway for ATP production, and is the preferred metabolic pathway utilized by cells with high energy demands . ConversBesides cellular energy and ATP production, intermediates produced from the different metabolic pathways, such as glycolysis, the PPP and the TCA cycle are also necessary for fatty acid and amino acid synthesis. For example the PPP and TCA cycles generate NADPH and citrate for fatty acid synthesis, respectively . The uniListeria monocytogenes infection , Zhu and colleagues highlighted that TAMs are of heterogeneous origin . They foNumerous studies have suggested that the metabolic pathways used by macrophages regulate their immune function . MacrophMetabolites generated during macrophage activation can impact and shape the inflammatory immune response. For instance, succinate, a metabolic product of the citric acid cycle accumulates in inflammatory macrophages to drive HIF-1α stabilization and IL-1β production . This meDecades of work have focused on tumor-cell metabolism, but it was not until the last decade did we learn that the highly glycolytic nature of tumor cells results in a nutrient-limited microenvironment . Tumor-iHIF-1α over-expression in macrophages upregulates glycolytic genes, such as pyruvate dehydrogenase kinase 1 (Pdk1), phosphoglycerate kinase 1 (Pgk1), glucose transporter 1 (Glut1), glucokinase (Gck) and pyruvate kinase isozymes M2 (Pkm2) . This rein vivo and in vitro (During aerobic glycolysis (the Warburg effect), pyruvate is converted to lactic acid by lactate dehydrogenase A (LDHA). Lactic acid, an end product of both aerobic and anaerobic glycolysis, activates vascular endothelial growth factor (VEGF), transforming growth factor β (TGF-β), and HIF-1α in oxidative tumor cells . At the in vitro . These din vivo mouse models for subcutaneous Lewis Lung carcinoma, orthotopic breast cancer and spontaneous mammary tumor, they observed that REDD1, an inhibitor of mTORC1 is highly upregulated in TAMs, particularly those located in hypoxic regions. Depletion of REDD1 in these macrophages greatly enhanced their glycolysis and they competed with neighboring endothelial cells for glucose. The restriction in glucose availability for the endothelial cells, in turn, stabilizes the vascular network to inhibit metastasis. These findings are in contrast to those reported by our group, where inhibiting glycolysis in TAMs resulted in a decrease in their pro-metastatic phenotype  metabolism, giving rise to differential macrophage functions (L-Arg into nitric oxide (NO) and L-citrulline. On the other hand, lL-4-polarized macrophages up-regulate Arg1, which catalyzes the conversion of L-Arg to L-ornithine, and polyamine synthesis  is a key lipogenic enzyme that catalyzes the terminal steps of fatty acid biogenesis and confers a growth and survival advantage to cancer cells . Cancer The role that myeloid-derived suppressor cells (MDSCs) have in regulating tumor progression is well recognized . MDSCs rMDSCs are comprised of two major subsets: monocytic MDSCs (M-MDSCs) and granulocytic MDSC (G-MDSCs). Both subsets can suppress the cytotoxic activity of cytotoxic CD8+ T lymphocytes (CTLs) and natural killer (NK) cells . In miceOver the course of inflammation, neutrophils engage in various cell–cell interactions with other immune cells, such as macrophages, dendritic cells and lymphocytes . NeutropThe tumor microenvironment polarizes TAMs toward a pro-tumoral M2-like phenotype, capable of promoting epithelial-to-mesenchymal transition in early pancreatic initiation and development . SimilarIn a recent study, The high concentration of lactate produced when tumor cells undergo Warburg metabolism as well as the hypoxic tumor microenvironment promote tumor progression by modifying the anti-tumoral immune response through priming tumor-infiltrating neutrophils toward an immunosuppressive state resulting in the induction of MDSCs . In turnA study conducted by Corzo and colleagues showed that tumor-infiltrating MDSCs suppressed both antigen-specific and non-specific T-cell activity, which was accompanied by up-regulation of Arg1 and iNOS, and down-regulation of NADPH oxidase and reactive oxygen species (ROS) . The mecAnother key sensors of cellular energy metabolism is adenosine monophosphate-activated protein kinase (AMPK) . AMPK isTumor cells derived lactic acid  mediated by HIF-1α can acts as an immunosuppressive metabolite and direct differential myeloid cell functions such as M2-like polarization . NeutropBoth M-MDSCs and G-MDSCs in tumor-bearing mice exhibit higher rates of glycolysis compared to their normal mature cell counterparts in healthy mice. By up-regulating glycolytic genes in response to tumor-derived factors and down-regulating ROS production to protect from apoptosis, MDSCs can accumulate in tumors . PhosphoThe glucocorticoid receptor (GR) is expressed by almost every cellular organism and is involved in regulating the genes that control energy metabolism and the immune response . GR signL-Arg depletion by G-MDSCs blocks CD3zeta expression in T cells resulting in the inhibition of antigen-specific proliferation , which has shown some therapeutic potential  and M2-like (MHCIIlow) TAMs are poor antigen presenters and are ineffective at stimulating naïve T-cell proliferation  suppressing polyclonal T-cell proliferation and (2) promoting an immunosuppressive T-cell phenotype in the tumor. Macrophages are the most abundant population in the tumor microenvironment, and have a crucial role in regulating the T-cell response. Unfortunately, both M1-like  are immunosuppressive cells that are critical for maintaining self-tolerance, especially in autoimmune diseases. In the context of cancer, the presence of Treg is often a negative prognostic factor with respect to overall survival  located around tumor/connective tissue also strongly secrete IL-10 and express more PD-L1 compared to other TAM subsets found within the tumor  , MJE3 E, 3-3-py, 3-dihydin vitro and in vivo studies reported encouraging results showing the ability of 3-BP to inhibit tumor growth (In vitro studies using mouse/rat hepatocytes (in vitro data clearly showed that 3-BP can impact on healthy tissues, particularly those that rely heavily on glycolysis. As such, using the same glycolytic inhibitors to target myeloid cells will also encounter the exact same adverse effects. Therefore, delivery of drugs specifically to the tumor will be the game-changer for these glycolytic drugs to progress form bench to bedside.The ideal anti-cancer therapy would be to specifically eradicate tumor cells with minimal effect on non-neoplastic populations in the body. This feature is even more critical for glycolytic inhibitors since glycolysis is a metabolic pathway utilized by almost all mammalian cells. For example 3-BP, the most “promising” anti-cancer drug mentioned earlier, was associated with causing death in at least three cancer patients . Early ir growth , yet onlr growth  and metar growth  were peratocytes  and primatocytes  reportedatocytes . These iOtto Warburg introduced the concept of Warburg metabolism, whereby tumor cells rely on glycolysis to support uncontrolled growth and proliferation . We now JS, SG, and SW conceptualized the content and wrote the manuscript. JS and SG prepared the figures.The 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.


Nintedanib is a tyrosine kinase inhibitor that efficiently slows the progression of idiopathic pulmonary fibrosis (IPF) and has an acceptable tolerability profile. In contrast, immune checkpoint inhibitors (ICIs) such as programmed death 1 and programmed death ligand 1 inhibitors have shown clinical activity and marked efficacy in the treatment of non-small cell lung cancer. However, it is unclear whether nintedanib reduces the risk of ICI-induced pneumonitis in IPF.A 78-year-old man with squamous cell lung carcinoma in IPF underwent second-line treatment with pembrolizumab. He was diagnosed as having pembrolizumab-induced pneumonitis after two cycles. He was administered prednisolone (PSL) and then improved immediately. Thereafter, his lung cancer lesion enlarged despite treatment with TS-1. Atezolizumab was then administered as 4th-line chemotherapy, but he immediately developed atezolizumab-induced pneumonitis after 1 cycle. The re-escalated dosage of PSL improved the pneumonitis, and then nintedanib was started as additional therapy. Under careful observation with nintedanib, atezolizumab was re-administered on day 1 of an every-21-day cycle. After three cycles, it remained stable without exacerbation of drug-induced pneumonitis.This case indicates the possibility that the addition of nintedanib to ICI therapy might prevent drug-induced pneumonitis or acute exacerbation of IPF. However, whether anti-fibrotic agents such as nintedanib are actually effective in preventing ICI-induced pneumonitis in ILD remains unknown and additional research is greatly needed to identify effective therapies for ILD combined with lung cancer. The treatment of advanced non-small cell lung cancer (NSCLC) has evolved to include targeted therapy, immune checkpoint inhibitors (ICIs), and chemotherapy for selected patients in the first-line setting. Angiogenesis inhibitors have been used in combination with chemotherapy in the first-line and maintenance settings to provide improved progression-free survival, objective response rate, and overall survival in selected studies. A biologic rationale exists for combining anti-angiogenic agents with immunotherapy and targeted kinase inhibitors . ICIs aiNintedanib is a tyrosine kinase inhibitor that efficiently slows the progression of idiopathic pulmonary fibrosis (IPF) and has an acceptable tolerability profile . TreatmeWe present the case of a 78-year-old man, a former smoker, with squamous cell lung carcinoma. Clinical staging was stage IV [cT3N2M1c (ADR)]. He was simultaneously diagnosed as having interstitial pneumonia. Chest high-resolution computed tomography (CT) showed a mass lesion of the right upper lobe as the primary lung carcinoma that was surrounded by ground-glass opacities as carcinomatous lymphangiomatosis. Interstitial pneumonia, as indicated by a subpleural reticular shadow with traction bronchiectasis and bronchiolectasis predominantly in the lower lobes and without apparent honeycombing, was comparable with probable usual interstitial pneumonia pattern based on recent criteria   were increased, although the size of the lung cancer tumors was reduced after 2 cycles  and PSL (5 mg/day), atezolizumab was re-administered on day 1 of an every-21-day cycle. After three cycles, it remained stable without exacerbation of drug-induced pneumonitis. However, afterwards, he developed multiple brain metastases and carcinomatous meningitis. Because his therapy was converted to palliative care, he died one and a half months later due to carcinomatous meningitis.Our patient developed drug-induced pneumonitis from both pembrolizumab and atezolizumab. However, the use of nintedanib allowed us to re-administer atezolizumab without further recurrence of the drug-induced pneumonitis. We highlight the possibility of using nintedanib to prevent atezolizumab-induced pneumonitis in IPF combined with lung cancer. ICIs such as programmed death 1 (PD-1) and PD ligand 1 inhibitors have shown clinical activity and marked efficacy in the treatment of NSCLC . PembrolNintedanib has a number of clinical benefits in patients with IPF, such as reducing the decline of lung function and extending the time to AE . NintedaIn conclusion, our case indicates the possibility that ICIs combined with nintedanib might prevent drug-induced pneumonitis or AE-IPF. However, whether anti-fibrotic agents such as nintedanib are actually effective in the prevention of ICI-induced pneumonitis in ILD remains unknown, and additional research is greatly needed to identify effective therapies for ILD combined with lung cancer.


Glioma is the most commonly diagnosed malignant and aggressive brain cancer in adults. Traditional researches mainly explored the expression profile of glioma at cell-population level, but ignored the heterogeneity and interactions of among glioma cells.Here, we firstly analyzed the single-cell RNA-seq (scRNA-seq) data of 6341 glioma cells using manifold learning and identified neoplastic and healthy cells infiltrating in tumor microenvironment. We systematically revealed cell-to-cell interactions inside gliomas based on corresponding scRNA-seq and TCGA RNA-seq data.A total of 16 significantly correlated autocrine ligand-receptor signal pairs inside neoplastic cells were identified based on the scRNA-seq and TCGA data of glioma. Furthermore, we explored the intercellular communications between cancer stem-like cells (CSCs) and macrophages, and identified 66 ligand-receptor pairs, some of which could significantly affect prognostic outcomes. An efficient machine learning model was constructed to accurately predict the prognosis of glioma patients based on the ligand-receptor interactions.Collectively, our study not only reveals functionally important cell-to-cell interactions inside glioma, but also detects potentially prognostic markers for predicting the survival of glioma patients.The online version of this article (10.1186/s12964-019-0363-1) contains supplementary material, which is available to authorized users. Glioma is the most common primary central nervous system (CNS) tumor in adults, and is known for its high heterogeneity and poor clinical outcomes . TraditiTumor together with surrounding stromal cells and extracellular matrix (ECM) constitute a tumorous niche referred as the tumor microenvironment (TME) , which pHere, we first explored the cell types of glioma cells using manifold learning based on a large amount of scRNA-seq data. Then the autocrine interactions among neoplastic cells were analyzed. Furthermore, we investigated the gene expression profile of macrophages and CSCs in gliomas, and subsequently examined the crosstalk between the two kinds of cell types. In the end, we built a robust machine learning model to predict the survival risk of glioma patients based on the expression level of specific ligands and receptors.https://www.ncbi.nlm.nih.gov/geo/) with accession number GSE89567. Bulk RNA-seq data of glioma were downloaded from The Cancer Genome Atlas (TCGA) data portal (https://tcga-data.nci.nih.gov/). Our analysis mainly focused on low grade glioma (LGG) and glioblastoma (GBM).We downloaded the single-cell RNA-seq gene expression data of glioma from GEO  . We. We38]. To further explore the interactions between CSCs and macrophages, we analyzed how macrophages communicated with CSCs via ligand-receptor pairs. For the ligands and receptors separately highly expressed in macrophages and CSCs, we identified 24 ligand-receptor pairs that showed crosstalk from macrophages to stem-like cells Fig. . We founP-value <0.05). The high expression of SDC1 is crucial for promoting stem-like cell invasion and metastasis in cell-extracellular matrix adhesion . Mo. Mo40]. adhesion . C3 playadhesion . We obser growth . Our resTo predict the prognosis of glioma patients, we built a model based on the ligand-receptor interactions between macrophages and CSCs using advanced machine learning algorithm XGBoost . FirstlyP-value = 0), which further confirmed the performance of our robust model  . Strikindel Fig. . The expdel Fig. . Among tdel Fig. . In addiThe evolution of tumor is a complicate progress, which involves the interactions between neoplastic and non-neoplastic cells infiltrating in the tumor microenvironment. The advantages of scRNA-seq provide unprecedented opportunities to investigate the cell-to-cell communications inside tumor. In this study, we systematically explored the communication networks between different cell types in glioma and gained insights into tumor microenvironment.In the microenvironment of gliomas, different kinds of cells were infiltrating and macrophages were enriched in tumor core site. We revealed that tumor cells could accomplish some biological processes through autocrine and paracrine ways. For instance, the neurite outgrowth inhibitor (Nogo) of RTN4 could play a newfound role in carcinogenesis through AKT signaling pathway, and knockdown of RTN4 could postpone tumor proliferation in mice . We founIn the crosstalk inside gliomas, the ligand-receptor signal pairs were associated with various biological processes, some of which may influence tumor progression. For instance, IL-8 has been shown to be strongly correlated with brain cancer grades , 42, andSince the ligands and receptors identified by us were significantly associated with the clinical outcomes of glioma, we built a robust model to predict the survival risk of patients to help clinical decisions using the advanced machine learning algorithm. Although the outcomes of patients could relate to the factors of age and general health conditions, our model still achieved good performance. Accordingly, our identified possible interactions in gliomas could be useful for the treatment and drug design of glioma , 55.Taken together, our results provide a landscape of the autocrine interactions in glioma neoplastic cells as well as the crosstalk between macrophages and glioma stem-like cells, which may facilitate the identification of potential therapeutic targets for precision medicine.Glioma is known for its heterogeneity and worse prognostic outcomes. In this study, we firstly revealed the cell-to-cell interactions inside gliomas using manifold learning based on a large amount of scRNA-seq data. Further analyses of the scRNA-seq and TCGA RNA-seq data of glioma provide a landscape of the autocrine interactions in glioma neoplastic cells as well as the crosstalk between macrophages and glioma stem-like cells. Some ligands and receptors identified by us could significantly affect prognostic outcomes and function via different pathways. Our machine learning model is able to accurately predict the prognosis of glioma patients based on the ligand-receptor interactions.Additional file 1:Table S1. 16 autocrine ligand-receptor pairs with significant Spearman’s correlation coefficients higher than 0.4. the first time they are cited. Table S2. 90 genes associated with stemness in glioma. Figure S1. Spearman’s correlation coefficients of two ligand-receptor pairs (GDFR-GFRA1 and RTN4-CNTNAP1) in TGCA LGG dataset. Figure S2. Enriched pathways for ligands highly expressed in stem-like cells and receptors highly expressed in macrophages (Pathway commons). Figure S3. Kaplan-Meier survival analysis for ITGB3 in TCGA LGG dataset. Figure S4. Enriched pathways for ligands highly expressed in macrophages and receptors highly expressed in stem-like cells (Pathway commons). Figure S5. PCA of IDH-mutated and IDH-wildtype gliomas based on the genes of ligand-receptor pairs. Figure S6. Performance of model built by randomly selected genes. (DOCX 546 kb)


Anti-glomerular basement membrane (anti-GBM) disease is characterized by circulating IgG glomerular basement membrane antibodies and is clinically expressed as a rapidly progressive crescentic glomerulonephritis (GN), with 30–60% of patients also developing pulmonary hemorrhage. Classically, the renal biopsy shows glomerular crescent formation, bright linear staining of glomerular basement membranes (GBM) for IgG on direct immunofluorescence (IF), and the serologic presence of circulating anti-GBM antibodies. Recently, patients with linear IgG IF staining, undetectable circulating anti-GBM antibodies and glomerular changes atypical for anti-GBM disease have been described as “atypical anti-GBM disease”, with a distinctly more benign clinical course than typical anti-GBM disease. We present a case report of a patient with negative anti-GBM serology but positive linear IgG staining by IF, severe diffuse crescentic and endocapillary proliferative glomerulonephritis, and renal failure, complicated by severe pulmonary hemorrhage after immunosuppression, likely due to cytomegalovirus (CMV) pneumonitis.A 24-year-old man was admitted to hospital with hemoptysis and renal failure. Investigations for anti-GBM serology by addressable laser bead immunoassay (ALBIA) was negative for anti-GBM antibodies. Renal biopsy showed diffuse endocapillary proliferative glomerulonephritis with membranoproliferative features and diffuse circumferential crescents. Direct IF showed strong linear staining for IgG along GBMs. The patient’s hemoptysis improved with immunosuppression, but 1 month later he was readmitted with gross hemoptysis, which was refractory to further cyclophosphamide, plasma exchange and rituximab. Bronchoalveolar lavage (BAL) and blood work confirmed CMV pneumonitis, and the patient’s hemoptysis resolved with ganciclovir, though he became dialysis dependent.This case demonstrates an atypical presentation of anti-GBM disease with both crescents and endocapillary hypercellularity and negative serology. The patient is dialysis dependent, unlike most previously described patients with atypical anti-GBM disease. The course was complicated by CMV pneumonitis, which contributed to the severity of the pulmonary manifestations and added diagnostic difficulty. Anti-glomerular basement membrane (anti-GBM) disease is a rare disease, characterized by circulating IgG anti-GBM antibodies, a rapidly progressive crescentic glomerulonephritis, with 30–60% of patients also developing pulmonary hemorrhage due to cross-reactivity of antibodies with alveolar basement membranes , 2. The In most patients, the anti-GBM antibodies are directed against the NC1 domain of the alpha-3 chain of type IV collagen, resulting in activation of the complement cascade and tissue injury typical of a type 2 immune response , 5. LighTreatment of anti-GBM glomerulonephritis involves high dose steroids, and cyclophosphamide for inhibition of antibody production and rebound hypersynthesis, along with plasma exchange for removal of circulating antibodies. , 4 RituxWe present a case of a 24-year-old male with atypical anti-GBM disease, marked by undetectable anti-GBM antibodies and unusual biopsy findings, who developed a relapse of pulmonary hemorrhage. After lack of response to immunosuppression, the hemorrhage was found to be most likely due to a cytomegalovirus (CMV) pneumonitis.A 24-year-old male with a history of hypertension, hypothyroidism, morbid obesity, and significant smoking history (both cigarette and marijuana) presented to urgent care with a two-week history of progressive leg edema and hemoptysis. He was found to have nephrotic range proteinuria and acute renal failure with a serum creatinine of 346 μmol/L . Chest x-rays showed progressive worsening bilateral patchy opacities  infection. Results of the second bronchoscopy confirmed CMV and a viral load of > 10This is a case of a 24-year-old male with aggressive atypical anti-GBM disease who developed recurrent massive hemoptysis after initial apparent response to induction therapy with corticosteroids and cyclophosphamide, subsequently found to have CMV viremia and pneumonitis as the likely cause for his severe clinical deterioration. The atypical pathology with severe phenotype, undetectable anti-GBM serology, and presence of CMV pneumonitis are the three main highlights of this case.To the best of our knowledge, this is the first case of atypical anti-GBM disease with a diffuse crescentic and endocapillary proliferative phenotype. Typical anti-GBM disease is associated with diffuse crescent formation without endocapillary or mesangial proliferation. , 13. On There are several possible reasons why histologically confirmed anti-GBM disease may be serologically negative by commercial immunoassays. One possible explanation is that the target antigen may be different, and thus the anti-GBM antibody is not detected by conventional laboratory immunoassays. An alternate antigen is also postulated to trigger alternate pathogenic pathways that result in a different pathologic appearance to the glomeruli , 10.Standard anti-GBM enzyme-linked immunosorbent assays (ELISAs) are considered highly sensitive and specific. However, they are designed to detect IgG, and may give negative results when circulating anti-GBM antibodies are IgA class. In addition, standard anti-GBM assays may be less sensitive for certain IgG subclasses such as IgG4 , 12. A rOur chemiluminescence assay  used a fluorochrome-coupled monoclonal anti-human IgG as the secondary antibody which has been shown to bind all subclasses of IgG but has highest sensitivity for IgG1 and IgG3 (manufacturer’s communication). Our patient may therefore have had anti-GBM IgG4 at a level undetectable by the assay despite being pathogenic, although the limited capacity of IgG4 to bind complement and engage Fc receptors on neutrophils raises questions regarding their pathogenic potential .This is a possible reason why serum anti-GBM remained negative throughout this patient’s course. The severe phenotype in association with anti-GBM IgG4 is unexpected in view of previous reports of mild disease associated with this subclass , 19 . InThe atypical renal pathology in our case, along with the IgM lambda monoclonal peak raised the possibility of proliferative glomerulonephritis with monoclonal immunoglobulin deposits (PGNMID) as well as Randall type monoclonal immunoglobulin deposition disease (MIDD). PGNMID and MIDD were ruled out as EM did not show any immune complex-type dense deposits nor ‘powdery’ linear densities along the GBM , 22,and Donaghy and Rees  noted a Secondly, to our knowledge, there are only two case reports that highlight the scenario of presumed relapse of anti-GBM disease, later discovered to be CMV induced pulmonary hemorrhage , 15. ChaThis patient’s presentation with endocapillary hypercellularity, segmental membranoproliferative features, linear GBM staining for IgG by IF, and negative anti-GBM serology fits previous descriptions of “atypical” anti-GBM disease.  It is, h


Heatstroke model mice administered Texas Red-dextran showed leakage outside the brain vessel walls. In addition, trans-endothelial electrical resistance (TEER) value was significantly reduced in induced pluripotent stem (iPS) cell-derived brain microvascular endothelial cells under heat stress by reducing claudin-5 expression. In addition, our results showed that the expression level of P-glycoprotein (P-gp) was increased in iPS cell-derived brain microvascular endothelial cells under heat stress. Furthermore, serum from heatstroke model mice could impair the BBB integrity of iPS cell-derived brain microvascular endothelial cells. These results suggest that BBB integrity was affected by heat stress in vivo and in vitro and provide important insights into the development of new therapeutic strategies for heatstroke patients.The incidence of heatstroke has been increasing. Heatstroke has been shown to affect physiological barrier functions. However, there are few studies of the effect of heat stress on the blood-brain barrier (BBB) function. In this study, we investigated the influence of heat stress on brain microvascular endothelial cells  The blood-brain barrier (BBB) consists of brain microvascular endothelial cells that are surrounded and supported by astrocytes and pericytes. It plays critical roles in brain homeostasis and neural function by regulating the transfer of substances from the peripheral circulation into the brain. Brain microvascular endothelial cells show high trans-endothelial electrical resistance (TEER) and express selective influx and efflux transporters, for the import and export of molecules, respectively. BBB dysfunction is associated with many neurological diseases, such as multiple sclerosis and traumatic brain injury –3.in vitro human models are considered to be useful for examining detailed analysis of the molecular mechanisms of BBB disruption by heatstroke. Lippmann et al. recently developed methods to differentiate into brain microvascular endothelial cells from human pluripotent stem cells, such as embryonic stem (ES) cells and induced pluripotent stem (iPS) cells ), the resistance value of an empty filter coated with collagen IV and fibronectin was subtracted from each measurement.The TEER values of the human iPS cell-derived brain microvascular endothelial cells incubated on the 24-well Transwell inserts were measured using Millicell ERS-2 (Merk-Millipore). To calculate the TEER  supplemented with HEPES (Sigma) and D-glucose (DPBS-H). DPBS-H containing 10 μg/mL sodium fluorescein  was then loaded into the upper chamber, and DPBS-H was added to the lower chamber. After incubation for 30 min, the medium in the lower chamber was collected, and the NaF concentration in the sample was determined using a fluorescence multi-well plate reader . The permeability coefficient was calculated as previously reported .Cells were cultured with 10% volume alamarBlue reagents (ThermoFisher Scientific). After incubation for 2 h, the medium was collected, and the cell viability was determined by an absorbance microplate reader .GAPDH). The sequences of the primers used in this study are listed in Total RNA was isolated using RNAiso Plus reagent (Takara), and the cDNA was synthesized with the SuperScript VILO cDNA synthesis kit (ThermoFisher Scientific). The levels of claudin-5, occludin, Zonula occludens-1 (ZO-1), multidrug resistance protein 1 , breast cancer resistance protein , and P-glycoprotein  mRNA were quantified using the SYBR Green detection system (Applied-Biosystems). The results were normalized to the housekeeping gene, glyceraldehyde 3-phosphate dehydrogenase  or a HRP-conjugated anti-rabbit IgG  at room temperature for 1 h. The bands were detected with ECL Plus western blotting detection reagents (ThermoFisher Scientific), and the signals were visualized with a LAS-4000 imaging system (Fuji Film).Cells were washed three times with sterile PBS, and cell lysates were obtained using radioimmunoprecipitation assay (RIPA) buffer (ThermoFisher Scientific) plus protease inhibitor cocktail (Roche). The proteins in the lysates were separated by SDS-PAGE on a 5–20% polyacrylamide gel (Wako) and were then transferred to polyvinylidene fluoride membranes (Merk-Millipore). After blocking with 5% skim milk in TBS containing 0.1% Tween 20 at room temperature for 1 h, the membranes were incubated with a mouse anti-claudin-5 antibody , a mouse anti-P-gp antibody , a rabbit anti-CD31 antibody , or a mouse anti-ß-actin antibody  at 4 Blood samples were collected from the inferior vena cava after exposure to either normal conditions  or heatstroke conditions  for 2 h, and the mice were sacrificed. Blood samples on ice for 2–3 h, then it was centrifuged, and the serum was collected. iPS cell-derived microvascular endothelial cells on Transwell membranes were treated with 10% serum from normal or heatstroke model mice in hESFM supplemented with 1% PDS and bFGF. After incubation for 24 h, the TEER values of the human iPS cells-derived brain microvascular endothelial cells on 24-well Transwell inserts were measured using Millicell ERS-2.Statistical analyses were performed using an unpaired two-tailed Student’s t-test.First, we evaluated the degree of BBB disruption in the heatstroke model mice. When Texas Red-dextran was intravenously administered to mice housed at normal temperature, it stayed within the circulation and did not leak into the brain  left. InLippmann et al. previously established a method for the differentiation of brain microvascular endothelial cells from human iPS cells [Next, we examined BBB permeability under heat stress using iPS cell-derived brain microvascular endothelial cells. PS cells , 11. TheBecause the paracellular permeability of microvascular endothelial cells is regulated by tight junctions, we analyzed the expression of the tight junction-related genes, including claudin-5, occludin, and ZO-1, in iPS cell-derived brain microvascular endothelial cells. The mRNA expression levels of occludin and ZO-1 were unchanged under heat stress . In contTo protect the brain from exposure to unnecessary metabolites and drugs and to provide nutrients to the brain, various efflux and influx transporters are expressed in brain microvascular endothelial cells . We nextIn Figs in vivo and in vitro using a mouse model and human iPS cell-derived brain microvascular endothelial cells. Previous studies have shown that hypoxia inducible factor-1α  and high-mobility group box 1 (HMGB1) were released from the gut lumen into the systemic circulation under heatstroke conditions . HMGB1  and LPS In summary, we showed that heat stress could induce BBB disruption by reducing claudin-5 expression. We also found that the expression of P-gp, which is a multidrug efflux transporter, could be regulated by heat stress in iPS cell-derived brain microvascular endothelial cells. Furthermore, our results showed that molecules that could disrupt BBB integrity were present in serum from heatstroke model mice. These results showed that heat stress might directly and indirectly regulate BBB function.


Right-sided heart failure is the major cause of death among patients who suffer from various forms of pulmonary hypertension and congenital heart disease. The right ventricle (RV) and left ventricle (LV) originate from different progenitor cells and function against very different blood pressures. However, differences between the RV and LV formed after birth have not been well defined. Work from our laboratory and others has accumulated evidence that redox signaling, oxidative stress and antioxidant regulation are important components that define the RV/LV differences. The present article summarizes the progress in understanding the roles of redox biology in the RV chamber-specificity. Understanding the mechanisms of RV/LV differences should help develop selective therapeutic strategies to help patients who are susceptible to and suffering from right-sided heart failure. Modulations of redox biology may provide effective therapeutic avenues for these conditions. Patients with pulmonary hypertension and repaired congenital heart disease are at risk for developing right-sided heart failure ,2,3. HowDevelopmentally, LV and RV myocytes are derived from different precursor cells. Cells in the first heart field (primary heart field) contribute to the formation of the LV myocardium, whereas cells in the second heart field (anterior heart field) construct the RV myocardium ,8,9. FunS-glutathionylation, nitrotyrosine formation, and S-nitrosocysteine formation were found to be higher in the failing RV compared to control RV [S-nitrotyrosinylated proteins include heat shock protein-90 and sarcoplasmic reticulum Ca2+-ATPase, and S-glutathionylated proteins include heat shock protein-90 and NADH-ubiquinone oxidoreductase [In rats, the injection of an inhibitor of the vascular endothelial growth factor receptor SU5416) plus stimuli such as hypoxia and ovalbumin immunization trigger pulmonary arterial hypertension ,15 and r plus stintrol RV . Mass speductase .Total protein carbonylation was not altered in the RVs of rats with pulmonary hypertension compared with the controls . HoweverGata4 promoter [GATA4 gene expression [GATA4 gene transcription is through releasing the negative regulation by annexin A1 [GATA4 gene expression -3-hydroxy-8′-apocarotenol (R)-3-hydroxy-8′-apocarotenol has potential to confer a similar activity as β-carotene. In addition, it possesses a hydroxyl group that may be redox active. Coprocholic acid and 1alpha,25-dihydroxy-25,25-diphenyl-26,27-dinorvitamin D3 also contain four and three hydroxyl groups, respectively.To investigate the possible differences between the adult RV and LV, metabolomics analysis was performed using RV and LV free wall tissues obtained from adult rats by examining molecules with mass less than 1000. While a majority of metabolites seem to be expressed at similar levels between the RV and LV, some molecules exhibited statistically significant difference between their levels in the two ventricles. From these data, four biologically relevant molecules that occur at different levels between the RV and LV were identified. Among them, the levels of (3arotenol A and coparotenol B were foarotenol C and neuarotenol D were foNeuromedin N and another larger peptide, neuromedin N, both elicit cell signaling by activating neurotensin receptors. Neuromedin N is a small peptide with six amino acids, Lys-Ile-Pro-Tyr-Ile-Leu, while neurotensin contains 13 amino acids, Glu-Leu-Tyr-Glu-Asn-Lys-Pro-Arg-Arg-Pro-Tyr-Ile-Leu. Interestingly, both peptides are relatively rich in carbonylation-susceptible amino acids described above. 33% of amino acids in neuromedin N and 38% in neurotensin are composed of carbonylation-susceptible amino acids.To investigate the possible role of neurotensin/neuromedin N in right heart failure, immunohistochemistry experiments using the neurotensin/neuromedin N antibody were performed to compare the RVs of control rats to those of rats with right heart failure promoted by the injection of SU5416 plus chronic hypoxia that promotes severe pulmonary hypertension . The exaStudies of experimental animals and human patients revealed that the RV is more susceptible to the occurrence of oxidative stress than the LV in the setting of heart failure because the RV has weaker antioxidant-adaptive defense mechanisms ,29,30.In female Wistar rats, Schreckenberg et al.  found thIn human heart failure patients, Borchi et al.  reported2 saturations.In children with tetralogy of Fallot, Chaturvedi et al.  observedThe RV health is crucial to patients with pulmonary hypertension as well as with congenital heart disease including tetralogy of Fallot, pulmonary atresia, transposition of the great arteries and hypoplastic left heart syndrome. While the global RV and LV gene expression patterns and cell signaling mechanisms are similar, there are some subtle but crucial differences between RV and LV that may potentially define chamber-specific therapeutic strategies. Redox signaling, oxidative stress status and antioxidant regulation define important RV/LV differences. Further work focusing on these biological mechanisms may improve preventative and/or treatment strategies to help patients who are susceptible to developing right-sided heart failure. We hope that this article will promote further basic research concerning redox mechanisms of RV failure and possibly future clinical trials for the use of redox agents for the treatment of right-sided heart failure.


Approximately 60% of those with dementia do not carry a diagnosis, undermining patient participation in clinical trials and family access to support. Under-diagnosis is driven by lack of knowledge about dementia, stigma, clinician inexperience and therapeutic nihilism. To address clinician-based contributors to under-diagnosis, we developed and implemented “Dementia 360,” a telementoring program modeled on the ECHO  framework. Remote participants (n=67) learned about the diagnostic process, pharmacological management, family support and dementia-related resources. The video-conference-based one-hour sessions occurred weekly over 2 months. Instruction was provided by a multi-disciplinary faculty team with extensive clinical experience. Didactic presentations were followed by case studies offered by participants. Physicians, nurses, psychologists and social workers from 40 organizations participated, of which 62% were from medically underserved rural clinics. Participants were administered pre- and post-program questionnaires about their level of confidence in assessing and treating individuals with memory loss and dementia-related behavioral symptoms. Of the 54 clinicians who completed pre-intervention confidence assessments, 30 completed post-assessment. The clinicians had significantly increased confidence in diagnosing and treating dementia and managing behavioral symptoms of dementia (p ranging from .0002 to .003). Qualitative feedback from focus groups was generally positive, for example, “Knowing the diagnosis criteria and steps to take to rule out other diagnoses will help me more accurately diagnose and rule out dementia for my patients.” Our findings suggest that delivering case-based education via ECHO has potential to increase clinician workforce confidence in diagnosing and managing dementia.


Recent studies have shown that TF-based genetic engineering could be a useful approach for improving plant abiotic stress tolerance because of the role of TFs as master regulators of clusters of stress-responsive genes. Here, we explore the use of abiotic stress-responsive TFs from CAM plants to improve abiotic stress tolerance and WUE in crops by controlling the expression of gene cohorts that mediate drought-responsive adaptations. Recent research has revealed several TF families including AP2/ERF, MYB, WRKY, NAC, NF-Y, and bZIP that might regulate water-deficit stress responses and CAM in the inducible CAM plant Mesembryanthemum crystallinum under water-deficit stress-induced CAM and in the obligate CAM plant Kalanchoe fedtschenkoi. Overexpression of genes from these families in Arabidopsis thaliana can improve abiotic stress tolerance in A. thaliana in some instances. Therefore, we propose that TF-based genetic engineering with a small number of CAM abiotic stress-responsive TFs will be a promising strategy for improving abiotic stress tolerance and WUE in crop plants in a projected hotter and drier landscape in the 21st-century and beyond.This perspective paper explores the utilization of abiotic stress-responsive transcription factors (TFs) from crassulacean acid metabolism (CAM) plants to improve abiotic stress tolerance in crop plants. CAM is a specialized type of photosynthetic adaptation that enhances water-use efficiency (WUE) by shifting CO Triticum aestivum L.) and maize (Zea mays L.) yields by 20.6% and 39.3%, respectively. High salinity, another abiotic stress that is harmful to crop production, affects 20% of total cultivated and 33% of irrigated agricultural lands worldwide  on marginal lands are already underway  to use for bioengineering abiotic stress tolerance in crop plants . For exaic lines .2 uptake and fixation to the nighttime, when evaporative water losses are minimal, and perform C3 carboxylation reactions when stomata are closed during the daytime. This temporal separation of carbon fixation leads to the formation of the four-carbon organic acid malate, which is stored in the vacuole during the night and subsequently undergoes decarboxylation to release CO2 for re-fixation during the day to produce carbohydrates  and UV-B irradiation levels than can agronomically important C3 plants -dependent and -independent response pathways . ABA-deptallinum , suggesttallinum .3 plants , Agave, and Opuntia as agricultural crops to provide food, forage, fiber, and biofuels  for CAM plants are now available for Agave  TF whose transcripts are 20-fold more abundant in CAM-performing older leaf pairs relative to C3-performing younger leaves in K. fedtschenkoi . Results from four, third-generation (T3) transgenic lines carrying the KlMYB59 indicate increased rosette size and biomass at 4-week-old juvenile stage, and increased shoot length at 8-week-old mature stage in transgenic plants compared to WT (Arabidopsis) were also more highly expressed in CAM-performing leaves of K. laxiflora and K. fedtschenkoi relative to C3-performing leaves . The function of this K. fedtschenkoi NAC83 TF (KfNAC83) is not known in CAM- or C3-performing CAM plants, but its A. thaliana ortholog suggests roles in abiotic stress responses and development (KfNAC83 shows enhanced water-deficit stress tolerance and increased integrated WUE in four independent transgenic T3-generation A. thaliana lines compared to WT plants (KfNAC83-overexpressing lines show significantly increased rosette size, leaves in the mature rosette, shoot biomass, number of siliques, and lateral roots compared to WT. Interestingly, these transgenic lines also showed tolerance to 150 mM NaCl. Collectively, our characterization results strongly suggest that at least two of these candidate CAM TFs have functions in abiotic stress responses and CAM photosynthesis.One of these candidate CAM TFs is a myeloblastosis (MYB59, closest ortholog in elopment . Functio3 photosynthesis plants. Our recent findings strongly suggest that a bioengineering approach using CAM abiotic stress-responsive TFs has the potential to increase abiotic stress tolerance in A. thaliana and possibly in C3 crop plants. Our results indicate that CAM abiotic stress-responsive gene expression can be modulated by the appropriate CAM TFs to generate stress-adaptive phenotypes in A. thaliana and likely other C3 plants because these CAM abiotic stress-responsive genes are apparently conserved and present in C3 plants (K. fedtschenkoi is distantly related to A. thaliana, transgenic A. thaliana lines carrying the obligate CAM plants’ TFs showed favorable features for translational applications. We are optimistic that overexpressing TFs from the inducible CAM halophyte, M. crystallinum will have similar favorable responses in A. thaliana lines. Co-overexpression of only a small number of obligate and/or inducible CAM plant abiotic stress-responsive TFs with demonstrated abiotic stress-adaptive or -responsive functions would provide a facile approach for bioengineering desirable responses to abiotic stress (The abiotic stress-adaptive features of CAM plants provide a wealth of genetic resources, specifically TFs, that are now available for functional testing and possible improvement of WUE and abiotic stress responses in C3 plants . Furtherc stress . Such an3 crop plant abiotic stress responses. AA, KR, and BWMW wrote the manuscript. AA, KR, and BW conducted the experiments and provided the CAM plant TF overexpression data. WY, TG, and JC provided the CAM plant TF sequences. All authors reviewed the final manuscript.BWMW conceived the CAM abiotic stress-responsive TF-based approach to improve CThe 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.