BackgroundSevere combined immunodeficiency (SCID) is one of the most severe forms of primary immunodeficiency. JAK3 gene is a critical determinant of SCID, as JAK3-STAT pathway regulates development, proliferation, activation, and differentiation of immune cells. This study aimed to identify the genetic cause of a family with a suspected SCID patient, and to perform carrier screening for two couples to assess the risk of conceiving offspring with birth defects.MethodsWhole-Exome Sequencing was performed on five individuals from the three families. A series of in vitro functional experiments, including Western blotting and luciferase assays, were conducted to assess the pathogenicity of the identified JAK3 variants.ResultsWe identified seven JAK3 variants, including five variants of uncertain significance (p.Arg402His, p.ILe688Phe, p.Leu129Phe, p.Met235Thr, p.Ala634Pro) and one pathogenic variant and one likely pathogenic variant (p.Gln1007Ter and p.Cys376Leufs*34). Among these, four variants (p.Gln1007Ter, p.Leu129Phe, p.Cys376Leufs*34 and p.Ala634Pro) were novel. In vitro functional experiments revealed that three of five variants of uncertain significance (VUSs) significantly reduced STAT5 phosphorylation and transcriptional activity, thereby reclassifying two variants (p.Arg402His and p.ILe688Phe) as likely pathogenic variants (LP) and one variant (p.Leu129Phe) as VUS with a Bayesian score of 3. In contrast, the remaining two variants (p.Ala634Pro and p.Met235Thr) did not affect JAK3 function, and were reclassified as VUS with a Bayesian score of 1 or 0.ConclusionThis study identified seven JAK3 variants from three families, including four novel variants. Functional experiments revealed that two VUSs were reclassified as LP and one VUS were reclassified as VUS with a Bayesian score of 3. These findings highlight the importance of integrating genetic and functional analyses to enhance diagnostic accuracy, inform treatment strategies for patients, clarify of the risk for carrier-screening couples, improve genetic counseling, and guide reproductive interventions.

Non-obstructive azoospermia (NOA) and cryptozoospermia are two significant conditions contributing to male infertility. However, the underlying genetic factors in most cases remain unknown. In our study, whole exome sequencing identified novel biallelic variants in TBC1 domain family member 8 (TBC1D8) in two patients. Patient P1 with NOA harbored c.890C>T (p.A297V) and c.2461G>A (p.V821I), and patient P2 with cryptozoospermia carried c.854C>T (p.P285L) and c.1912G>A (p.D638N). Bioinformatic analyses predicted that all identified TBC1D8 variants were likely pathogenic. Compared with a normal control, patient P1 showed reduced expression of TBC1D8 in testicular tissue. Subsequently, hematoxylin-eosin staining and immunofluorescence analysis of testicular sections showed defective acrosome formation and the absence of elongated spermatids in patient P1, resulting from abnormal autophagy. Additionally, intracytoplasmic sperm injection treatment was beneficial for patient P2 with cryptozoospermia. In conclusion, our results suggest that TBC1D8 is a potentially novel candidate gene for male infertility associated with NOA or cryptozoospermia in humans.

Nasopharyngeal carcinoma (NPC) is prevalent in East and Southeast Asia, with genetic factors playing a significant role in its occurrence. The HLA gene region on chromosome 6 is linked to NPC susceptibility, but the mechanisms remain unclear. Epstein-Barr virus (EBV) infection is a well-established cause, with 95% of NPC patients being EBV-positive. Three key variations in the EBV genome (162215_C, 162476_C, 163364_T) in the BALF2 gene are strongly associated with NPC risk. This study finds that the high-risk BALF2 variant (BALF2-HR) upregulates HLA class II molecules, such as HLA-DP, which interacts with LAG-3 on CD8⁺ T cells, inhibiting cytokine secretion and promoting T cell exhaustion, leading to immune evasion and reduced anti-PD-1 efficacy. BALF2-HR also enhances HLA-DP transcription by binding to KPNA2 and facilitating CIITA nuclear translocation. Conjunctive immunotherapy with anti-LAG-3 and anti-PD-1 antibodies significantly improves NPC treatment. This work introduces a new therapeutic strategy for NPC and insights into infection-associated cancers.

INTRODUCTION:FHND-9041 is a novel third-generation EGFR tyrosine kinase inhibitor (EGFR-TKI). This first-in-human study is a single-arm, multi-center, open-label, non-randomized phase Ⅰ/II trial that aims to evaluate the tolerability, safety, pharmacokinetics, and anti-tumor activity of FHND-9041 in patients with EGFR-mutated non-small cell lung cancer (NSCLC). METHODS:The phase I study enrolled 87 patients with previously-treated EGFR T790M-positive NSCLC. The dose escalation study was conducted at doses of 40, 80, 120, and 180 mg/day, with an expansion of 36 and 39 patients in the 80 mg and 120 mg dose groups, respectively, to evaluate safety and pharmacokinetics. With 80 mg QD as the recommended phase II dose, the phase II study assessed the efficacy and safety of FHND-9041 as first-line treatment in 37 treatment-naive patients. RESULTS:No dose-limiting toxicity was observed, and the maximum tolerated dose was not identified within the 40 - 180 mg dose range. Pharmacokinetic data indicated dose-proportional exposure up to 120 mg, with a plateau at 120 mg. The phase II study revealed an objective response rate of 62.2 % (23/37 evaluable patients) with a median progression-free survival of 15.5 months (95 % CI, 12.9 - 18.1). The most common adverse events were decreased white blood cell count (22.6 %) and diarrhea (21.8 %). Treatment-related grade 3/4 adverse events included decreased white blood cell count (11.3 %) and liver impairment (4.8 %). CONCLUSIONS:FHND-9041 exhibits favorable safety and efficacy profile, supporting its further clinical development for EGFR-mutated advanced NSCLC.

Regulatory T cells (Treg cells) have reshaped modern immunology by establishing the conceptual and mechanistic foundation of peripheral immune tolerance. Since the pioneering identification of CD4(+)CD25(+) suppressive T cells by Shimon Sakaguchi and the subsequent discovery of the lineage-defining transcription factor forkhead box P3 (Foxp3) by Mary E. Brunkow and Fred Ramsdell, Treg cells have been recognized as indispensable guardians of immune homeostasis. These advances collectively clarified that central tolerance alone is insufficient to eliminate all self-reactive lymphocytes, and peripheral tolerance-critically mediated by Treg cells-serves as a second barrier preventing pathological autoimmunity. Contemporary research has therefore expanded the functional and therapeutic significance of Treg cells across the fields of autoimmunity, cancer, transplantation, and tissue repair. Treg cells originate from two major developmental pathways: thymus-derived Treg (tTreg) cells, which arise from high-affinity self-reactive TCR interactions in the thymus, and peripheral Treg (pTreg) cells, which are induced in mucosal and other peripheral tissues via antigen stimulation under tolerogenic cytokine cues such as IL-2 and TGF-beta. Their differentiation is orchestrated by a multilayered transcriptional and epigenetic network within the Foxp3 locus, including CNS0-CNS3 elements that integrate TCR, cytokine and environmental signals to support lineage stability. Treg cells are identified by a combination of surface and intracellular markers-CD25, CD127(low/-),CTLA-4, GITR, TNFR2, CD39/CD73, and Foxp3-although marker specificity varies with context, activation state, and species. Their notable heterogeneity enables Treg cells to adopt Th1-, Th2-, Th17-or Tfh-like programs through transcription factors such as T-bet, GATA3, ROR gamma t and Bcl6, thereby permitting precise suppression of corresponding effector responses. Tissue-resident Treg subsets in adipose tissue, skin, skeletal muscle and the CNS have emerged as highly specialized regulators that integrate local metabolic and stromal signals, contributing not only to immunosuppression but also to tissue regeneration. Mechanistically, Treg cells maintain tolerance through three synergistic strategies: (1) secretion of suppressive cytokines (IL-10, TGF-beta, IL-35) and cytotoxic mediators (granzyme B, perforin); (2) cell-contact-dependent interactions via CTLA-4, PD-1/PD-L1, and LAG-3 to limit dendritic cell maturation and T-cell activation; and (3) metabolic regulation including IL-2 consumption, adenosine production via CD39/CD73, cAMP transfer through gap junctions, and adaptation to hypoxic or nutrient-restricted microenvironments. Dysregulation of Treg cell quantity or function contributes directly to pathogenesis across a spectrum of diseases. In autoimmune diseases such as type 1 diabetes, systemic lupus erythematosus, rheumatoid arthritis and multiple sclerosis, impaired Foxp3 stability, epigenetic abnormalities, defective IL-2 signaling or inflammatory cytokine exposure undermine Treg suppressive capacity, facilitating excessive autoreactive T-and B-cell activation. In contrast, within the tumor microenvironment, Treg cells are often enriched through chemokine axes such as CCL22-CCR4 and reinforced by interaction with myeloid-derived suppressor cells and tumor-associated macrophages. Their enhanced metabolic fitness and suppressive phenotype enable tumors to evade immune destruction. In transplantation, Treg cells are essential for promoting graft tolerance, restraining effector T-cell activation, and facilitating tissue repair after injury. Rapid therapeutic progress has been driven by Treg-based immunomodulation. Polyclonal Treg adoptive transfer has demonstrated safety and preliminary efficacy in type 1 diabetes, autoimmune disorders, solid-organ transplantation, and graft-versus-host disease. Gene-engineered Treg therapies, including antigen-specific CAR-Treg and TCR-Treg platforms, offer superior precision and stability, enabling targeted suppression at disease sites. Additional strategies-including low-dose IL-2 therapy, small-molecule modulation, and selective depletion of intratumoral Treg using antibodies against CCR4, CCR8, CTLA-4 or CD25xTIGIT bispecifics-further expand the translational landscape. Collectively, advances in Treg biology-from lineage ontogeny and molecular regulation to specialized functions and therapeutic engineering -highlight Treg cells as central orchestrators of immune equilibrium. Continued integration of single-cell multi-omics, systems immunology and gene-editing technologies is expected to accelerate the development of highly specific, durable and safe Treg-centered therapies, ultimately enabling precision control of immune tolerance in autoimmunity, transplantation and cancer.

Immune evasion represents a crucial milestone in the progression of cancer and serves as the theoretical foundation for tumor immunotherapy. In this study, we reveal a negative association between Human Papillomavirus (HPV)-encoded circular RNA, circE7, and the infiltration of CD8+ T cells in head and neck squamous cell carcinoma (HNSCC). Both in vitro and in vivo experiments demonstrate that circE7 suppresses the function and activity of T cells by downregulating the transcription of LGALS9, which encodes the galectin-9 protein. The molecular mechanism involves circE7 binding to acetyl-CoA carboxylase 1 (ACC1), promoting its dephosphorylation and thereby activating ACC1. Activated ACC1 reduces H3K27 acetylation at the LGALS9 gene promoter, leading to decreased galectin-9 expression. Notably, galectin-9 interacts with immune checkpoint molecules TIM-3 and PD-1, inhibiting the secretion of cytotoxic cytokines by T cells and promoting T cell apoptosis. Here, we demonstrate a mechanism by which HPV promotes immune evasion in HNSCC through a circE7-driven epigenetic modification and propose a potential immunotherapy strategy for HNSCC that involves the combined use of anti-PD-1 and anti-TIM-3 inhibitors.

Breast cancer is the most prevalent malignancy and the most significant contributor to mortality in female oncology patients. Potassium Two Pore Domain Channel Subfamily K Member 1 (KCNK1) is differentially expressed in a variety of tumors, but the mechanism of its function in breast cancer is unknown. In this study, we found for the first time that KCNK1 was significantly up-regulated in human breast cancer and was correlated with poor prognosis in breast cancer patients. KCNK1 promoted breast cancer proliferation, invasion, and metastasis in vitro and vivo. Further studies unexpectedly revealed that KCNK1 increased the glycolysis and lactate production in breast cancer cells by binding to and activating lactate dehydrogenase A (LDHA), which promoted histones lysine lactylation to induce the expression of a series of downstream genes and LDHA itself. Notably, increased expression of LDHA served as a vicious positive feedback to reduce tumor cell stiffness and adhesion, which eventually resulted in the proliferation, invasion, and metastasis of breast cancer. In conclusion, our results suggest that KCNK1 may serve as a potential breast cancer biomarker, and deeper insight into the cancer-promoting mechanism of KCNK1 may uncover a novel therapeutic target for breast cancer treatment.

BACKGROUND:N6-methyladenosine (m6A) modification is essential for modulating RNA processing as well as expression, particularly in the context of malignant tumour progression. However, the exploration of m6A modification in nasopharyngeal carcinoma (NPC) remains very limited. METHODS:RNA m6A levels were analysed in NPC using m6A dot blot assay. The expression level of methyltransferase-like 14 (METTL14) within NPC tissues was analysed from public databases as well as RT-qPCR and immunohistochemistry. The influences on METTL14 expression on NPC proliferation and metastasis were explored via in vitro as well as in vivo functional assays. Targeted genes of METTL14 were screened using the m6A and gene expression profiling microarray data. Actinomycin D treatment and polysome analysis were used to detect the half-life and translational efficiency of ANKRD22. Flow cytometry, immunofluorescence and immunoprecipitation were used to validate the role of ANKRD22 on lipid metabolism in NPC cells. ChIP-qPCR analysis of H3K27AC signalling near the promoters of METTL14, GINS3, POLE2, PLEK2 and FERMT1 genes. RESULTS:We revealed METTL14, in NPC, correlating with poor patient prognosis. In vitro and in vivo assays indicated METTL14 actively promoted NPC cells proliferation and metastasis. METTL14 catalysed m6A modification on ANKRD22 messenger ribonucleic acid (mRNA), recognized by the reader IGF2BP2, leading to increased mRNA stability and higher translational efficiency. Moreover, ANKRD22, a metabolism-related protein on mitochondria, interacted with SLC25A1 to enhance citrate transport, elevating intracellular acetyl-CoA content. This dual impact of ANKRD22 promoted lipid metabolism reprogramming and cellular lipid synthesis while upregulating the expression of genes associated with the cell cycle (GINS3 and POLE2) and the cytoskeleton (PLEK2 and FERMT1) through heightened epigenetic histone acetylation levels in the nucleus. Intriguingly, our findings highlighted elevated ANKRD22-mediated histone H3 lysine 27 acetylation (H3K27AC) signals near the METTL14 promoter, which contributes to a positive feedback loop perpetuating malignant progression in NPC. CONCLUSIONS:The identified METTL14-ANKRD22-SLC25A1 axis emerges as a promising therapeutic target for NPC, and also these molecules may serve as novel diagnostic biomarkers.

Abstract In South and Southeast Asia, the habit of chewing betel nuts is prevalent, which leads to oral submucous fibrosis (OSF). OSF is a well‐established precancerous lesion, and a portion of OSF cases eventually progress to oral squamous cell carcinoma (OSCC). However, the specific molecular mechanisms underlying the malignant transformation of OSCC from OSF are poorly understood. In this study, the leading‐edge techniques of Spatial Transcriptomics (ST) and Spatial Metabolomics (SM) are integrated to obtain spatial location information of cancer cells, fibroblasts, and immune cells, as well as the transcriptomic and metabolomic landscapes in OSF‐derived OSCC tissues. This work reveals for the first time that some OSF‐derived OSCC cells undergo partial epithelial–mesenchymal transition (pEMT) within the in situ carcinoma (ISC) region, eventually acquiring fibroblast‐like phenotypes and participating in collagen deposition. Complex interactions among epithelial cells, fibroblasts, and immune cells in the tumor microenvironment are demonstrated. Most importantly, significant metabolic reprogramming in OSF‐derived OSCC, including abnormal polyamine metabolism, potentially playing a pivotal role in promoting tumorigenesis and immune evasion is discovered. The ST and SM data in this study shed new light on deciphering the mechanisms of OSF‐derived OSCC. The work also offers invaluable clues for the prevention and treatment of OSCC.

Transfer RNAs (tRNAs) play pivotal roles in the transmission of genetic information, and abnormality of tRNAs directly leads to translation disorders and causes diseases, including cancer. The complex modifications enable tRNA to execute its delicate biological function. Alteration of appropriate modifications may affect the stability of tRNA, impair its ability to carry amino acids, and disrupt the pairing between anticodons and codons. Studies confirmed that dysregulation of tRNA modifications plays an important role in carcinogenesis. Furthermore, when the stability of tRNA is impaired, tRNAs are cleaved into small tRNA fragments (tRFs) by specific RNases. Though tRFs have been found to play vital regulatory roles in tumorigenesis, its formation process is far from clear. Understanding improper tRNA modifications and abnormal formation of tRFs in cancer is conducive to uncovering the role of metabolic process of tRNA under pathological conditions, which may open up new avenues for cancer prevention and treatment.

Clinicopathological data for 72 NPC tissues and 20 non-cancerous NPE samples from patients with chronic nasopharyngeal inflammation using RT-PCR.

Currently, more than 170 modifications have been identified on RNA. Among these RNA modifications, various methylations account for two-thirds of total cases and exist on almost all RNAs. Roles of RNA modifications in cancer are garnering increasing interest. The research on m 6 A RNA methylation in cancer is in full swing at present. However, there are still many other popular RNA modifications involved in the regulation of gene expression post-transcriptionally besides m 6 A RNA methylation. In this review, we focus on several important RNA modifications including m 1 A, m 5 C, m 7 G, 2′-O-Me, Ψ and A-to-I editing in cancer, which will provide a new perspective on tumourigenesis by peeking into the complex regulatory network of epigenetic RNA modifications, transcript processing, and protein translation.

Mass spectrometry identification of proteins pulled down by biotin-labeled circBART2.2 probe.

Single cell RNA sequencing (scRNA-seq) provides a great convenience for studying tumor occurrence and development for its ability to study gene expression at the individual cell level. However, patient-derived tumor tissues are composed of multiple types of cells including tumor cells and adjacent non-malignant cells such as stromal cells and immune cells. The spatial locations of various cells in situ tissues plays a pivotal role in the occurrence and development of tumors, which cannot be elucidated by scRNA-seq alone. Spatially resolved transcriptomics (SRT) technology emerges timely to explore the unrecognized relationship between the spatial background of a particular cell and its functions, and is increasingly used in cancer research. This review provides a systematic overview of the SRT technologies that are developed, in particular the more widely used cutting-edge SRT technologies based on next-generation sequencing (NGS). In addition, the main achievements by SRT technologies in precisely unveiling the underappreciated spatial locations on gene expression and cell function with unprecedented high-resolution in cancer research are emphasized, with the aim of developing more effective clinical therapeutics oriented to a deeper understanding of the interaction between tumor cells and surrounding non-malignant cells.

List of antibodies for immunohistochemistry, western blotting, immunofluorescence, flow cytometry, RNA pulldown, and ChIP experiments.

为了适应临床医学八年制医学生的培养目标,推进医学免疫学教学改革,中南大学基础医学院免疫学系探索构建线下一流课程教学模式,在充分理解医学免疫学教学内容特点基础上,对每堂课课前、课中和课后三阶段进行整体设计,采用多元化的教学方法,融合多种教学改革举措,通过基础-临床融通改善教学效果,结合课程思政,以提高医学免疫学教学质量.文章结合课程设计、实践以及教师的体会,总结该线下课程教学模式的方法和特点,以期为培育创新型人才为导向的医学教育以及打造其他学科的线下一流课程提供经验.

Immune evasion represents a crucial milestone in the progression of cancer and serves as the theoretical foundation for tumor immunotherapy. Currently, immune checkpoint inhibitors such as anti-PD-1/PD-L1 have shown promising therapeutic efficacy in clinical settings for various tumors. However, a significant proportion of tumor patients fail to benefit from this class of immune checkpoint inhibitors, implying potential involvement of other immune checkpoint molecules in tumor immune escape. In this study, we unveiled a negative association between Human Papillomavirus (HPV)-encoded circular RNA circE7 and the presence of infiltrating CD8+ T cells in head and neck squamous cell carcinoma (HNSCC). Both in vitro and in vivo experiments demonstrated that circE7 suppressed the function and activity of T cells by downregulating the transcription of LGALS9, which encodes the galectin-9 protein. The unexpected molecular mechanism involved the binding of circE7 to acetyl-CoA carboxylase 1 (ACC1), enhancing its dephosphorylation, and thereby activation of ACC1, which resulted in a reduction in the acetylation level of H3K27 in the promoter region of LGALS9 gene and subsequent downregulation of galectin-9 expression. Notably, galectin-9 interacts with immune checkpoint molecules TIM-3 and PD-1 present on T cell surface, triggering T cells to secrete cytotoxic cytokines and hindering T cell apoptosis. In essence, HPV-mediated downregulation of galectin-9 expression through circE7 impairs T cell function and activity, ultimately promoting immune evasion in HNSCC. Importantly, we confirmed that a combination of monoclonal antibodies simultaneously targeting TIM-3 and PD-1 significantly improved the immunotherapeutic efficacy in HNSCC both in vitro and in animal models. Our study unveils a novel mechanism through which HPV promotes immune evasion in HNSCC via circE7-mediated epigenetic modification, offering a new therapeutic strategy for enhanced HNSCC immunotherapy.

Probes for fluorescence in situ hybridization and RNA pulldown, siRNAs, primers for ChIP assays and RT-PCR, and luciferase reporter vectors.

The 2023 Nobel Prize in Physiology or Medicine was awarded to medical scientists Katalin Kariko and Drew Weissman for their discovery of nucleoside base modification. The two researchers found that mRNA produced from modified nucleoside bases can evade innate immune recognition and improve protein expression. The discovery of these modifications is essential for the development of an effective mRNA vaccine against novel coronavirus SARS-CoV-2. Vaccination is the most economical and effective measure to prevent infectious diseases. So far, the vaccine has entered the third generation of nucleic acid vaccine from inactivated vaccine, subunit vaccine and recombinant protein vaccine. mRNA vaccine has the potential of high efficiency, rapid development, low-cost production and safe use. It is not a replication vector, and it does not have the characteristics of antibiotic resistance, genome integration and strong immunogenicity. Furthermore, corresponding mRNA vaccine can be quickly produced for different strains in large-scale, which speeds up the production process. However, mRNA synthesized in vitro is the ligand of Toll-like receptors. Once mRNA synthesized in vitro activates these receptors, the innate immune responses in the body will be turned on and a large amount of type I interferon will be produced, and mRNA will face the risk of degradation. The study of Katalin Kariko and Drew Weissman found that in vitro transcribed mRNA with modified bases can escape poor immune activation and solve the problem of inflammatory response stimulated by in vitro transcribed mRNA. Further studies have found that mRNA containing pseudouridine can be translated more effectively. At the same time, Drew Weissman has made an important contribution to the research and development of delivery system. After the outbreak of COVID-19, based on the research of two scientists, together with the development of efficient delivery systems such as lipid nanoparticles (LNPs) delivery system and one-component ionizable amphiphilic Janus dendrimer delivery system, the stability of SARS-CoV-2 prickle antigen and unparalleled investment by industry and government, the research and development technology system of mRNA vaccine was improved. Two very successful mRNA-based SARS-CoV-2 vaccines were approved at the end of 2020, which played a very important role in fighting COVID-19 during COVID-19 epidemic and paved the way for future mRNA applications. mRNA vaccine also has great potential in inhibiting tumor growth and regenerative medicine. Therefore, the circular RNA vaccine can also be studied. This paper introduces the process of vaccine development, the important nucleotide modification and lipid nanoparticles technology in mRNA vaccine, the in vivo effect of mRNA vaccine and the summary and prospect of technical development.