Research
Abstract
Ex vivo engineering strategies for adoptive αβ T-cell therapies increasingly use pharmacological modulation to improve survival, expansion, and antitumor activity. Short-term exposure to the BCL-2 inhibitor venetoclax during αβ T-cell manufacturing enhances apoptotic priming and effector persistence, suggesting a route to strengthen other T-cell lineages. γδ T cells share cytotoxic properties with αβ T cells but recognize targets independently of major histocompatibility complex (MHC) and show low alloreactivity, supporting off-the-shelf use in acute myeloid leukemia (AML). Whether such conditioning benefits γδ T cells was unknown. Here, we show that ex vivo venetoclax pretreatment enhances the antileukemic efficacy of therapeutic γδ T cells and chimeric antigen receptor (CAR) γδ T cells. Venetoclax-pretreated γδ T cells displayed increased cytotoxicity and proliferation with reduced exhaustion, yielding superior control of AML blasts and xenografts. These functional gains coincided with elevated mitochondrial content and a fatty acid oxidation metabolic profile. In vivo, venetoclax-pretreated γδ T cells achieved durable disease suppression, and the same conditioning improved CAR γδ T-cell efficacy. Together, these results show that short-term BCL-2 inhibition enhances γδ T-cell cytotoxicity and persistence. Incorporating venetoclax pretreatment into γδ T-cell manufacturing may improve therapeutic efficacy and inform next-generation γδ T-cell therapies for AML.
Authors
Xingchi Chen, Lin Zhang, Bingbing Yan, Yinqiang Sui, Weiwei Ma, Hui Zhao, Yining Wang, Kepeng Yang, Jiewen Ma, Baolin Tang, Yonghui Zhang, Xiaoyu Zhu
Abstract
Regulatory T (Treg) cells in visceral adipose tissue (VAT) play essential roles in systemic metabolic homeostasis under distinct physiological and pathological conditions. However, the metabolic cues that drive Treg cell subset specialization in the obese VAT niche remain elusive. Here, we demonstrated that palmitic acid instigated chronic VAT inflammation and systemic metabolic disturbance by compromising the immunosuppressive function of the ICOShi Treg subset. Palmitic acid, but not oleic acid, activated Crebzf expression in VAT Treg cells from HFHS diet-induced obese and ob/ob mice. Crebzf deficiency significantly attenuated diet-induced obesity and inflammation by upregulating the suppressive function of VAT ICOShi Treg cells. Moreover, adoptive transfer of Crebzf-deficient ICOShi Treg cells into Rag1-/- mice alleviated HFHS diet-induced inflammation and metabolic disorders more effectively than transfer of Crebzf-sufficient ICOShi Treg cells. Mechanistically, CREBZF interacted with c-JUN to inhibit Foxp3 activity, thereby impairing the stability and inhibitory cytokine production of ICOShi Treg cells. In human subjects, CREBZF levels in VAT Treg cells were elevated and negatively correlated with FOXP3 activity. Collectively, these findings uncover a specific ICOShi Treg subset that responds to palmitic acid, thereby coupling obesogenic signals to VAT remodeling and systemic metabolic homeostasis.
Authors
Weitong Su, Yuxiao Liu, Xi Yan, Mengyao Huang, Linghao Xu, Jing Lin, Xufeng Chen, Puyuan Hu, Chenlin Gao, Jian Wen, Hongdong Wang, Dong Ding, Zengpeng Zheng, Wenjing Li, Lianjia Li, Zhan Liu, Keyu Qian, Jing Gao, Tingting Zhang, Xiaobing Mao, Haibing Zhang, Wei Lu, Bin Li, Hong Li, Aoyuan Cui, Yan Bi, Chunxiang Zhang, Yu Li
Abstract
Human carcinomas often gain aggressive characteristics and escape cell-type specific treatment regimens through cryptic shifts in lineage states. However, the underlying mechanisms that govern lineage plasticity in carcinomas are undefined. Here in this study, we found that PAX5, a neural/lymphatic transcription factor, contributed to neuroendocrine (NE) lineage transition. PAX5 was highly expressed in aggressive human NE carcinoma cells and tissues but not in non-NE cancer cells and tissues. Deletion of Pax5 in Rb1fl/fl;Trp53fl/fl mice caused a reduction of tumor vessels, loss of NE morphologic features and decreased expression of ASCL1, NCAM, and SYP, whereas ectopic expression of PAX5 in CC10-rtTA;TetO-hEGFRex19del/T790M mice adenocarcinomas and in LNCAP prostate cancer xenografts induces an angiogenic microenvironment and NE morphology. Importantly, antiangiogenic drugs reduced NE features of Rb1fl/fl;Trp53fl/fl tumors and blocked PAX5-induced NE transformation. These studies demonstrate an essential role of angiogenic microenvironment in transition/maintenance of NE lineage, suggesting that targeting PAX5 and its downstream signaling may modulate lineage transitions responsible for treatment failure in both SCNCs and adenocarcinomas.
Authors
Ailing Wu, Yujie Hao, Xuemiao Yan, Junrong Liu, Lin Wang, Yan Jin, Wenxu Liu, Xiyue Chen, Yuan Jiang, Luc Girard, Zhiqun Shang, Jun Yan, Zhenfa Zhang, Wenchen Gong, Yuanjie Niu, Benjamin J. Drapkin, John D. Minna, Lance S. Terada, Zhenyi Ma, Zhe Liu
Abstract
Chronic primary pain conditions (CPPCs), such as fibromyalgia and vestibulodynia, affect over 100 million Americans, predominantly women, and pose a substantial healthcare challenge. CPPCs arise from genetic and environmental factors that enhance catecholamine tone, potentially through miRNA dysregulation following catecholamine activation of beta-adrenergic receptors. Here, we identified miR-133a-3p as a biomarker of CPPC status and investigated its functions using in vivo and in vitro approaches. Plasma levels of miR-133a-3p were consistently downregulated in humans with ≥1 CPPC and in rat and mouse models of primary pain. Our data suggest that miR-133a-3p is packaged in extracellular vesicles that are secreted by adipocytes and trafficked to the spinal cord. Activation of adrenergic receptors on white adipocytes resulted in downregulation of miR-133a-3p which negatively regulated pain-related genes in the spinal cord, such as MAP3K3, which is critical for sensory neuron activation. Adipose-specific overexpression of miR-133a-3p in a mouse model of primary pain reversed mechanical hypersensitivity in both sexes. These findings implicate miR-133a-3p dysregulation in primary pain across conditions and species and establish its role in multi-site mechanical hypersensitivity. Further, miR-133a-3p overexpression shows therapeutic potential for the millions of individuals with CPPCs.
Authors
Nathaniel P. Hernandez, Jiegen Chen, Yiling Qian, Xin Zhang, Yaomin Wang, Brittney P. Ciszek, Xianglong Gao, Marguerita E. Klein, Yun-Ling Pai, Mohamad Karaky, Carolina B. Meloto, Francesca Montagna, Matt Kanke, Clair Crewe, Luda Diatchenko, Praveen Sethupathy, Andrea G. Nackley
Abstract
Neddylation is highly activated in many human cancers and may serve as a therapeutic target for clinical treatment. However, it remains unclear regarding the role of neddylation in tumor angiogenesis. Here, we demonstrate that the neddylation E2 enzyme UBE2M is upregulated in tip cells and is essential for tumor vascular sprouting. We show that UBE2M-mediated neddylation of STAT1 enhances its phosphorylation and promotes the transcription of DLL4. This elevated DLL4 expression in tip cells activates Notch signaling in adjacent stalk cells, thereby maintaining the tip-stalk cell balance and ensuring organized vascular patterning. Consequently, endothelial-specific deletion of UBE2M reduces DLL4 expression, leading to excessive but non-productive sprouting due to uncontrolled tip cell formation and lack of stalk cell support, which ultimately suppresses tumor growth. Importantly, targeting endothelial neddylation potently sensitizes various tumors to anti-VEGF therapy. Together, our findings unveil UBE2M as a key regulator of angiogenic signaling and identify it as a promising anti-angiogenic target in cancer.
Authors
Xinyi Jiang, Jie Zhang, Li Zhou, Zonglin Li, Ningcong Sun, Xian Xu, Jisong Zhang, Yizhou Huang, Xue Zhang, Enguo Chen, Hongqiang Cheng, Yuehai Ke
Abstract
Regulatory T cells (Tregs) maintain immune tolerance through mechanisms tightly coupled to cellular metabolism. Whereas glycolysis supports Treg migration, lipid metabolism sustains their suppressive phenotype. Here, we identify the sterol regulatory element–binding protein 1c (SREBP1c) as a central regulator of Treg immunobiology. Tregs from Srebp1c-deficient mice displayed impaired suppressive function, reduced frequencies in circulation and lymphoid tissues, and diminished expression of functional markers. These defects stemmed from intrinsic metabolic rewiring rather than systemic alterations, as both ex vivo Tregs (CD4+CD25hiFoxP3+) and in vitro-derived Tregs lacking Srebp1c were shifted toward glycolysis. Integrated transcriptomic and lipidomic analyses revealed that Srebp1c-deficient Tregs exhibited defective phospholipid remodeling, with an accumulation of lysophosphatidylcholines over phosphatidylcholines, which we attributed to enhanced cytosolic phospholipase A2 (cPLA2α) activity and disruption of the Lands cycle. Altered lipid composition impaired adenosine-mediated immunosuppression by reducing CD73 expression and extracellular adenosine generation. Accordingly, pharmacological inhibition of cPLA2α restored adenosine signaling, CD73 expression, and Treg suppressive capacity. Thus, by preserving phospholipid homeostasis, SREBP1c functions as an immunometabolic checkpoint that links lipid metabolism to adenosine-dependent Treg suppression.
Authors
Fabrizia Bonacina, Claudio Procaccini, Marta Iaia, Arianna Moretti, Monika Svecla, Silvia Pedretti, Jeroen F.J. Bogie, Giovani Battista Vingiani, Annalisa Moregola, Francesca Genova, Claudia Russo, Giusy De Rosa, Claudia La Rocca, Giada Mondanelli, Marco Gargaro, Nico Mitro, Giuseppe Matarese, Giuseppe Danilo Norata
Abstract
Efferocytosis, the clearance of apoptotic cells by macrophages, promotes tissue resolution. Efficient resolution requires efferocytosis-induced macrophage proliferation (EIMP) to expand pro-resolving macrophages. Here, we show that efferocytosis activates base excision repair (BER) to remove 8-OHdG from DNA, enabling EIMP. Mechanistically, efferocytosis promotes poly(ADP-ribose) polymerase-1 (PARP1) chromatin binding and PARylation to facilitate DNA repair complex assembly, and increases nuclear MTH1/NUDT1, which hydrolyzes 8-OHdG. Both processes require DNA-methyltransferase-3A (DNMT3A), which is activated during efferocytosis. Using a model where dexamethasone-induced thymocyte apoptosis triggers efferocytosis-mediated thymic repair, we showed that DNMT3A is required for increases in nuclear PARP1/MTH1, oxidized DNA suppression, EIMP in thymic macrophages, and thymic repair. We next studied a human-relevant model of atherosclerosis regression, where efferocytosis drives protective lesional fibrous cap thickening. We compared WT mice with a model of DNMT3A-clonal hematopoiesis (CH), in which loss-of-function DNMT3A mutations promote atherosclerotic disease. Atherosclerosis regression in WT mice led to decreased nuclear 8-OHdG and increases in nuclear PARP1/MTH1 and EIMP in lesional macrophages and fibrous cap thickening, all of which were impaired in DNMT3A-CH regression. These findings reveal that efferocytosis initiates a BER pathway to allow macrophage proliferation for tissue resolution, with possible therapeutic relevance to atherosclerosis regression and DNMT3A-CH.
Authors
Kleopatra Avrampou, Santosh R. Sukka, David Ngai, Patrick Ampomah, Xiaobo Wang, George Kuriakose, Jacob Glass, Bernhard Dorweiler, Hanna Winter, Lars Maegdefessel, Hanrui Zhang, Aaron Viny, Ira Tabas
Abstract
Defective endometrial decidualization is one major cause of female infertility, yet the underlying mechanisms remain elusive. Here, we identified that protein arginine methyltransferase 5 (PRMT5) which was upregulated during decidualization and by progesterone stimulation, is markedly down-regulated in the endometria of patients with recurrent implantation failure (RIF), along with a global reduction of symmetric dimethylarginine (sDMA). Uterine stromal-specific ablation of Prmt5 in mouse severely impaired decidualization leading to infertility. A multi‑omics analysis in human endometrial stromal cells (EnSCs) revealed that PRMT5 promoted decidualization primarily by catalyzing sDMA at arginine 346 (R346) of the orphan nuclear receptor Nur77, which directs its proper chromatin occupancy. Targeting the PRMT5-Nur77 methylation axis, we designed a peptide, Pep‑Nur77-R346K, which rescued the decidualization of multiple preclinical models: PRMT5 deficient human EnSCs, both genetic knockout (Prmt5d/d) and pharmacologically inhibited mouse models, an estrogen deficient mouse model, and, more importantly, the primary RIF EnSCs. In a retrospective cohort of 114 participants, the correlated reductions of endometrial PRMT5/Nur77-R346me2s was confirmed, which demonstrated robust predictive value for pregnancy outcome. Our work establishes the PRMT5‑Nur77 methylation axis as a key regulator of endometrial receptivity, and highlights both a novel diagnostic biomarker and a peptide‑based therapeutic potential for fertility.
Authors
Zhiwen Cao, Xinyu Cai, Jie Mei, Na Kong, Yang Liu, Xiaoyue Shen, Min Wu, Xin Zhen, Jianxin Sun, Rong Li, Ruiwei Jiang, Haixiang Sun, Guijun Yan
Abstract
Epilepsy affects approximately 50 million people worldwide, yet more than half of individuals with a presumed genetic cause still lack a molecular diagnosis despite the identification of over 1,000 monogenic epilepsy genes. This diagnostic gap is unlikely to be resolved by improved variant detection alone, suggesting that variants affecting the same biological pathway may combine to cause disease. By studying epilepsy-associated actin regulatory genes, we identified a conserved “actin-mitochondria-glutamate (AMG) pathway”. We demonstrate that reduced actin polymerization promotes DRP1-mediated mitochondrial fission, increases reactive oxygen species (ROS) levels, and enhances glutamatergic transmission, leading to seizures. The glial innate immune pathway, a recently recognized contributor to epilepsy, is activated when the AMG pathway is affected. Reducing mitochondrial fission with the DRP1 inhibitor Mdivi-1, or suppressing ROS with N-acetyl-L-cysteine amide (NACA), significantly alleviates seizures. Importantly, digenic heterozygous loss‑of‑function variants in AMG‑pathway genes combine to cause seizures, and individuals with epilepsy of unknown etiology show an increased burden of such variants when compared to the controls. Modeling patient‑specific digenic combinations in Drosophila confirms that many combinations promote seizure susceptibility. Together, these findings establish the AMG pathway as a mechanistic framework for identifying digenic etiologies in epilepsy and highlight potential therapeutic targets.
Authors
Shenzhao Lu, Mengqi Ma, Shabab B. Hannan, Mingxi Deng, Hu Chen, Zhijian Yu, Lindsey D. Goodman, Haein Kim, Yun Zhao, Sandeep Kumar Dubey, Wen-Wen Lin, Xueyang Pan, Debdeep Dutta, Vishnu Anand Cuddapah, Jill A. Rosenfeld, Xi Luo, Zhandong Liu, Joshua M. Shulman, Hugo J. Bellen
Abstract
Hepatic stellate cell (HSC) activation can lead to liver fibrosis, for which there are no effective treatments. Aberrant cytoskeletal reorganization is a central driver of HSC activation. Non-muscle myosin II (NM II) is known to regulate cytoskeleton remodeling via its actin cross-linking and contractile properties. However, the molecular players controlling actomyosin assembly and contractility in HSCs during liver fibrosis remain poorly defined. Here, we identified integrin β-like 1 (ITGBL1) as a gatekeeper of HSC quiescence by negatively regulating actomyosin contractility-driven mechanotransduction in HSCs. ITGBL1 expression was markedly elevated in activated HSCs found in patient and mouse fibrotic livers. Unexpectedly, HSC-specific Itgbl1 deficiency worsened liver fibrosis, whereas ITGBL1 overexpression in HSCs limited it, suggesting a protective role for ITGBL1 against a pathogenic HSC activation. Multi-omics and functional analyses revealed that ITGBL1 impaired F-actin filament organization in HSCs by disrupting myosin heavy chain 9 (MYH9, also named NM II heavy chain A)-dependent actomyosin assembly. In line, HSC-specific Myh9 deficiency or silencing of Myh9 in HSCs alleviated liver fibrosis. Taken together, our findings unveil the ITGBL1-MYH9 interaction acts as a critical mechano-regulatory brake that maintains cytoskeletal equilibrium and mechanical homeostasis in HSCs, providing a promising therapeutic strategy to combat liver fibrosis.
Authors
Yixin Li, Yan Wang, Chenhao Tong, Xinghuan Fu, Ningning Ma, Yawen Hao, Zian Feng, Shijia Ling, Zequn Yin, Haodong Li, Shujun Ge, Siting Yang, Peng Xiao, Siyue Dong, Adrien Guillot, Yajun Duan, Yong He
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Copyright © 2026 American Society for Clinical Investigation
ISSN: 0021-9738 (print), 1558-8238 (online)