Introduction:Neoantigens from the Kirsten rat sarcoma viral oncogene homolog (KRAS) are specific cancer therapeutic targets. However, to date, no immune product targeting KRAS neoantigens has been approved for clinical use, and key challenges regarding efficacy and generalizability remain. Methods:In this study, we isolated a natural human T-cell antigen receptor (TCR) 0 that specifically recognized human leukocyte antigen (HLA)-A*11:01+ T2 cells pulsed with KRAS G12V8-16 peptides. However, TCR0 gene-transduced T cells demonstrated inadequate response to tumor cell lines. We generated T cells expressing a TCR0 mutant, being designated as TCR3. Results:TCR3-T cells showed significantly optimized avidity and response to tumor cell lines, retained specificity for the KRAS G12V8-16 peptide with no response to normal cells, killed tumor cells that highly expressed programmed cell death-ligand 1 in vitro and in vivo, proliferated without being seriously affected by indoleamine 2,3-dioxygenase, resisted transforming growth factor β, and infiltrated and recruited other immune cells to the tumor site through chemokines. Discussion:TCR3 may be useful for KRAS neoantigen-targeted clinical immunotherapy, help resolve cancer immune escape, and enhance clinical effectiveness and safety.

BACKGROUND:CD8⁺ T cell exhaustion is a defining feature of the immunosuppressive TME in ccRCC. METHODS:We employed a multi-omics driven pipeline to nominate Nicotinamide N-methyltransferase (NNMT) as a high-confidence therapeutic target in ccRCC. This computational prediction was validated through bulk RNA-seq, single-cell RNA sequencing, and spatial transcriptomics to delineate NNMT-associated molecular and cellular programs. While the discovery phase highlighted endothelial-specific NNMT overexpression, we further validated the functional consequences of NNMT modulation using Caki-1 and A498 cell lines to model the downstream signaling cascades. Functional assays assessed impacts on proliferation, apoptosis, cytokine secretion (IL-6, IL-1β, TNF-α), and TGF-β pathway activity. Immune infiltration and T cell exhaustion signatures were evaluated across TCGA cohorts. RESULTS:Multi-omics profiling revealed that NNMT is specifically overexpressed in tumor-associated endothelial cells enriched for active TGF-β signaling and inflammatory cues. High NNMT expression strongly correlated with CD8⁺ T cell exhaustion, elevated apoptotic signaling, and immunosuppressive cytokine production. In functional validation, NNMT knockdown suppressed TGF-β activity, reduced pro-inflammatory cytokines, and restored CD8⁺ T cell infiltration and effector function. Mechanistically, NNMT loss shifted the BAX/Bcl-2 ratio toward apoptosis and increased cleaved caspase-3. Spatial transcriptomics confirmed that NNMT⁺ endothelial cells form an immunosuppressive niche in direct contact with exhausted T cells. We also found that I-BET-762, I-BET-151, PFI-1, and BMS-387032 can target and inhibit NNMT to reduce CD8⁺ T cell exhaustion. CONCLUSION:We establish NNMT as a central metabolic-immune hub that orchestrates TGF-β-mediated CD8⁺ T cell dysfunction and endothelial reprogramming in ccRCC.

Abstract Background Worldwide, lung cancer is the most common cause of cancer-related deaths. Molecular targeted therapies and immunotherapies for non-small-cell lung cancer (NSCLC) have improved outcomes markedly over the past two decades. However, the vast majority of advanced NSCLCs become resistant to current treatments and eventually progress. A traditional Chinese medicine (TCM) formula of Shuangshen granules (SSG) has demonstrated potential in alleviating cancer side effects and improving survival rate. Despite clinical evidence supporting its benefit, there is still insufficient understanding of the active compounds in SSG and their underlying mechanisms, which limits its broader clinical application. Methods Lewis lung carcinoma (LLC) tumor-bearing mouse model was established to assess the efficacy of combined SSG and anti-PD-1 therapy in vivo, and myeloid-derived suppressor cells (MDSC) and CD8+T cells were isolated for in vitro co-culture experiments, while pathological examination was conducted using hematoxylin and eosin (HE). The expression of PD-1, TIM-3, CTLA-4, LAG-3, Arg-1, IDO, iNOS, PD-L1 and Gal-9 was detected using immunohistochemistry (IHC), immunofluorescence, and flow cytometry and Western blotting. The expression of IL-2, TNF-α and IFN-γ were detected by reverse transcription-quantitative polymerase chain reaction (qPCR). Concentrations of IL-10 and TGF-β were measured by enzyme-linked immunosorbent assay (ELISA). Network pharmacology and molecular docking were utilized to screen for potential therapeutic targets and intervening signaling pathways of SSG in lung adenocarcinoma (LUAD). The predictions derived from this approach were further verified using Western blotting. Results In vivo experiments using LLC xenograft mice demonstrated that SSG suppressed tumor growth in a dose-dependent manner, with high-dose SSG showing optimal efficacy in inhibiting tumor angiogenesis and cell proliferation. SSG enhances anti-tumor immunity by reducing T cell exhaustion and MDSC-mediated immunosuppression, with SSG + anti-PD-1 combination therapy synergistically optimizing the tumor immune microenvironment. Network pharmacology analysis revealed 5 hub targets (IL2, STAT3, HSP90AA1, LGALS3, and FGF2) associated with immune, LUAD, and active ingredients of SSG, with significant enrichment in the PI3K-Akt pathway. Compared with the control group, the protein expression levels of p-PI3K and p-Akt in the SSG group were significantly down-regulated, indicating that the PI3K-Akt pathway may be inhibited. Conclusions SSG could dose-dependently inhibit LLC tumor growth in mice and exert antitumor effects by alleviating T-cell exhaustion and MDSC-mediated immunosuppression. Notably, IL2, STAT3, HSP90AA1, LGALS3 and FGF2, as potential targets of SSG, were significantly enriched in the PI3K-Akt pathway, which provide a novel perspective for the treatment of LUAD.

Traditional aging clocks, typically taking chronological age or mortality as targets, face a trade-off between predictive accuracy and biological interpretability. To address this limitation, we developed protein organ clocks (POA), a novel approach that integrates aging indices derived from organ function indicators and blood proteomic data. Systematic comparison with four traditional blood protein clocks and three organ-specific clocks revealed that POA outperformed these models in associating with aging markers, predicting the risk of major organ-related diseases and mortality, and identifying clinically relevant organ aging subtypes, capturing inter-individual heterogeneity in organ aging. Critically, we constructed reduced POA models using 387 proteins that closely replicate the performance of full models, significantly lowering analytical burden and enabling feasible large-scale clinical application. The superior performance of POA stems from its integrated analysis of organ function and plasma proteomics, which enhances aging signal detection, improves biological interpretability, and strengthens clinical translational potential. To facilitate broad adoption, we developed a web server (https://www.biosino.org/poa) that allows users to upload proteomics data and calculate POAs for the body and eight organs. This work establishes a new paradigm for aging quantification and informs strategies to extend healthy lifespan and optimize disease prevention.

Lung cancer, predominantly non-small cell lung cancer (NSCLC), remains a principal driver of cancer-related morbidity and mortality worldwide. Despite advancements in surgery, radiotherapy, chemotherapy, and targeted treatments, outcomes remain poor in advanced NSCLC. The tumor microenvironment (TME) exerts a critical influence on therapy responses. Within the TME, immune cells such as T and B lymphocytes, dendritic cells, myeloid-derived suppressor cells, tumor-associated macrophages, neutrophils, and natural killer cells can drive both pro- and anti-tumor processes. This review integrates their classification, phenotypic plasticity, and roles in NSCLC, highlighting key preclinical and clinical evidence while discussing pathogenesis, prognostic significance, and therapeutic potential. We also summarize the current immunotherapeutic strategies for advanced NSCLC, including first- or second-line regimens with immune checkpoint inhibitors alone or combined with chemotherapy, anti-angiogenic agents, or additional checkpoint inhibitors, and future directions. By elucidating the interplay between the NSCLC immune microenvironment and emerging immunotherapies, this review emphasizes the need for novel combination regimens and robust predictive biomarkers to improve clinical outcomes and extend survival in advanced NSCLC.

Pulmonary hypertension is one of the most intractable cardiovascular diseases in the world today. The current pharmacological treatments can temporarily relieve symptoms, but the therapeutic effect is unsatisfactory in advanced pulmonary hypertension with severe pulmonary vascular remodeling and right ventricular hypertrophy. The nuclear receptor superfamily is a vital class of transcriptional regulators in the human body. Extensive research has demonstrated that these regulators play pivotal roles in a multitude of physiological processes and are involved in the regulation of the pathogenesis of a variety of pathological diseases, providing potential therapeutic targets for pulmonary hypertension. The targeting of nuclear receptors, such as MR, PPARγ, and NUR77, has demonstrated potential in improving pulmonary vascular remodeling and preventing the progression of experimental pulmonary hypertension. However, further in-depth mechanistic exploration and large-scale preclinical studies are necessary to facilitate eventual clinical application. In this review, we describe in detail the various types of nuclear receptors that affect the function of pulmonary vascular endothelial cells and smooth muscle cells by regulating inflammation, oxidative stress, metabolism, and other processes, thereby affecting the prognosis of pulmonary hypertension, and highlight the clinical therapeutic potential of nuclear receptors. Intensive research on the regulatory role of nuclear receptors in pulmonary hypertension has yielded new avenues for targeted therapy of pulmonary hypertension and has prompted the exploration of new therapeutic strategies to combat this life-threatening disease.

Cyclin-dependent kinase 4 and 6 inhibitors (CDK4/6 inhibitors) can significantly extend tumor response in patients with metastatic luminal A breast cancer, yet intrinsic and acquired resistance remains a prevalent issue. Understanding the molecular features of CDK4/6 inhibitor sensitivity and the potential efficacy of their combination with novel targeted cell death inducers may lead to improved patient outcomes. Herein, we demonstrate that ferroptosis, a form of regulated cell death driven by iron-dependent phospholipid peroxidation, partly underpins the efficacy of CDK4/6 inhibitors. Mechanistically, CDK4/6 inhibitors downregulate the cystine transporter SLC7A11 by inhibiting SP1 binding to the SLC7A11 promoter region. Furthermore, SLC7A11 is identified as critical for the intrinsic sensitivity of luminal A breast cancer to CDK4/6 inhibitors. Both genetic and pharmacological inhibition of SP1 or SLC7A11 enhances cell sensitivity to CDK4/6 inhibitors and synergistically inhibits luminal A breast cancer growth when combined with CDK4/6 inhibitors in vitro and in vivo. Our data highlight the potential of targeting SLC7A11 in combination with CDK4/6 inhibitors, supporting further investigation of combination therapy in luminal A breast cancer.

TSHR is a member of the glycoprotein hormone receptors, a subfamily of class A G-protein-coupled receptors and plays pivotal roles in various physiological and pathological processes, particularly in thyroid growth and hormone production. The aberrant TSHR function has been implicated in several human diseases including Graves' disease and orbitopathy, nonautoimmune hyperthyroidism, hypothyroidism, cancer, neurological disorders, and osteoporosis. Consequently, TSHR is recognized as an attractive therapeutic target, and targeting TSHR with small-molecule modulators including agonists, antagonists, and inverse agonists offers great potential for drug discovery. In this perspective, we summarize the structures and biological functions of TSHR as well as the recent advances in the development of small-molecule TSHR modulators, highlighting their chemotypes, mode of actions, structure-activity relationships, characterizations, in vitro/in vivo activities, and therapeutic potential. The challenges, new opportunities, and future directions in this area are also discussed.

Antigen-specific T cell receptor-engineered T cell (TCR-T) based immunotherapy has proven to be an effective method to combat cancer. In recent years, cross-talk between the innate and adaptive immune systems may be requisite to optimize sustained antigen-specific immunity, and the stimulator of interferon genes (STING) is a promising therapeutic target for cancer immunotherapy. The level of expression or presentation of antigen in tumor cells affects the recognition and killing of tumor cells by TCR-T. This study aimed at investigating the potential of innate immune stimulation of T cells and engineered T cells to enhance immunotherapy for low-expression antigen cancer cells. We systematically investigated the function and mechanism of cross-talk between STING agonist diABZI and adaptive immune systems. We established NY-ESO-1 full knockout Mel526 cells for this research and found that diABZI activated STING media and TCR signaling pathways. In addition, the results of flow cytometry showed that antigens presentation from cancer cells induced by STING agonist diABZI also improved the affinity of TCR-T cells function against tumor cells in vitro and in vivo. Our findings revealed that diABZI enhanced the immunotherapy efficacy of TCR-T by activating STING media and TCR signaling pathways, improving interferon-γ expression, and increasing antigens presentation of tumor cells. This indicates that STING agonist could be used as a strategy to promote TCR-T cancer immunotherapy.

Interface residues at sites of protein-protein interaction (PPI) are the focus for affinity optimisation. However, protein hydrophobic cores (HCs) play critical roles and shape the protein surface. We hypothesise that manipulating protein HCs can enhance PPI interaction affinities. A cell stress molecule, major histocompatibility complex class I chain-related protein A (MICA), binds to the natural killer group 2D (NKG2D) homodimer to form three molecule interactions. MICA was used as a study subject to support our hypothesis. We redesigned MICA HCs by directed mutagenesis and isolated high-affinity variants through a newly designed partial-denature panning (PDP) method. A few mutations in MICA HCs increased the NKG2D-MICA interaction affinity by 325-5613-fold. Crystal structures of the NKG2D-MICA variant complexes indicated that mutagenesis of MICA HCs stabilised helical elements for decreasing intermolecular interactive free energy (ΔG) of the NKG2D-MICA heterotrimer. The repacking of MICA HC mutants maintained overall surface residues and the authentic binding specificity of MICA. In conclusion, this study provides a new method for MICA redesign and affinity optimisation through HC manipulation without mutating PPI interface residues. Our study introduces a novel approach to protein manipulation, potentially expanding the toolkit for protein affinity optimisation.

AbstractBackgroundAstragaloside IV (AS‐IV) is the most active monomer in the traditional Chinese herbal medicine Radix Astragali, which has a wide range of antiviral, anti‐inflammatory, and antifibrosis pharmacological effects, and shows protective effects in acute lung injury.MethodsThis study utilized the immunofluorescence, flow cytometry, enzyme‐linked immunosorbent assay, quantitative reverse transcription‐polymerase chain reaction, western blot, and hematoxylin and eosin staining methods to investigate the mechanism of AS‐IV in reducing viral pneumonia caused by influenza A virus in A549 cells and BALB/c mice.ResultsThe results showed that AS‐IV suppressed reactive oxygen species production in influenza virus‐infected A549 cells in a dose‐dependent manner, and subsequently inhibited the activation of nucleotide‐binding oligomerization domain‐like receptor thermal protein domain associated protein 3 inflammasome and Caspase‐1, decreased interleukin (IL) ‐1β and IL‐18 secretion. In BALB/c mice infected with Poly (I:C), oral administration of AS‐IV can significantly reduce Poly (I:C)‐induced acute pneumonia and lung pathological injury.ConclusionsAS‐IV alleviates the inflammatory response induced by influenza virus in vitro and lung flammation and structural damage caused by poly (I:C) in vivo.

Significance For over half a century, the three main pillars of conventional cancer therapy are surgery, chemotherapy, and radiotherapy. However, these treatment methods have inherent limitations, as they inevitably cause severe damage to normal cells, particularly immune cells. The discovery and development of immunotherapy show promising clinical applications. Nonetheless, immunotherapy is a double-edged sword, often leading to the occurrence of immune-related adverse events (irAEs) because of off-target effects. Therefore, the current focus in cancer research is to explore treatment strategies that can activate local immune responses while enhancing tumor specificity. Near-infrared photoimmunotherapy (NIR-PIT) is a novel tumor therapy, and it depends on a single antibody-photo absorber conjugate (APC), which combines a monoclonal antibody (McAb) targeted on tumor features with IRDye700DX (IR700). Except for its specific antitumor mechanisms, a unique aspect of NIR-PIT is its direct impact on blood drug delivery. The super-enhanced permeability and retention (SUPR) effects facilitate the rapid leakage of drugs into the tumor, favoring the induction of cytotoxic effects. However, the presence of the "binding site barrier" indicates that using antibodies with low affinity or targeting antibodies with low antigen expression may promote a more even distribution of APCs within the tumor parenchyma. In recent years,researchers have investigated the use of different targeting segments in NIR-PIT, enhancing the tumor immunogenicity, targeting ability, stability, and flexibility of NIR-PIT drugs. This approach has shown considerable potential for application in various types of tumors, with some related clinical trials yielding satisfactory results. Studies have shown a close association between the suppressive tumor microenvironment (TME) and the growth and progression of cancer. In recent years, the targets of APCs in NIR-PIT have expanded to surface proteins of non-tumor cells in TME. The combination therapy of NIR-PIT with immune checkpoint blockade (ICB) has also shown promising experimental results. The development and continuous improvement of optical devices also facilitate the monitoring and evaluation of the therapeutic effects of NIR-PIT. Therefore, it is necessary to summarize previous relevant research to provide a rational reference for the clinical research and application of NIR-PIT. Progress The primary mechanism through which NIR-PIT exerts its cytotoxic effects is via a photochemical reaction from IR700.Under near-infrared light, IR700 in APCs undergoes a photocatalytic transformation, changing its chemical properties from hydrophilic to hydrophobic, and aggregating in an aqueous solution. This process leads to the denaturation of the cell membrane antigens bound to it, physical damage to the cell membrane and cell rupture, increased transmembrane water flow, and cell death (Fig.1). Simultaneously, the rapid release of tumor-associated antigens (TAAs) and damage-associated molecular patterns (DAMPs)during NIR-PIT induces immunogenic cell death (ICD), and subsequently, activates the antitumor immune response of the host,enhancing the activation of systemic immune responses to attack other cancer cells and further amplifying the therapeutic effects of NIR-PIT (Fig.2). Current NIR-PIT treatment strategies targeting key components in TME, including immune inhibitory cells (Tregs and MDSCs), cancer-associated fibroblasts (CAFs), and blood vessels, are listed in Table 1. The design principles of APCs and relevant experimental results are also presented. Subsequently, the combined therapeutic strategies and efficacy of immune checkpoint inhibitors with NIR-PIT targeting different cell surface proteins are elucidated. Given the heterogeneity of immune cell populations in different tumors, the choice of ICBs can be based on the expression levels of specific immune checkpoint molecules in their respective tumors. In addition, the progress of clinical trials related to NIR-PIT is summarized, demonstrating that cetuximab-IR700(RM-1929) can elicit effective antitumor responses in patients with locally recurrent HNSCC where conventional clinical treatments are less effective. Furthermore, the SUPR effect can be quantified using indocyanine green (ICG)-fluorescence and magnetic resonance imaging (MRI) contrast agents to monitor and identify the viability of NIR-PIT. 18F-fluorode-oxyglucose positron emission tomography (18F-FDG-PET), fluorescence lifetime imaging, and bioluminescence imaging can evaluate acute NIR-PIT treatment in preclinical studies. Moreover, the micro distribution of the NIR-PIT agent and its therapeutic effects is monitored using a two-channel fluorescence fiber-imaging system and two-photon microscopy with and without a microprism. The tumoricidal effects and hemodynamic changes induced by NIR-PIT can be monitored by C-13 MRI, blood oxygenation level dependent (BOLD) MRI, and photoacoustic imaging. The current investigation of NIR-PIT is relatively limited. In summary, the limitations of replicating IRdye700-McAb conjugates in NIR-PIT, the penetration and uniformity of near-infrared light irradiation, differences in the types and expression of target molecules in different types of tumors, and the safe range of APC dosage in NIR-PIT still require detailed investigations. Conclusions and ProspectsExtensive in vitro and in vivo models study on NIR-PIT have been conducted for various types of tumors, with promising therapeutic outcomes. With its broad and flexible application scope, and various approaches to enhance its efficacy, NIR-PIT has significant potential as a valuable method for cancer treatment.

Abstract Immune checkpoint inhibitors (ICIs) has revolutionized the treatment landscape of various cancers by reinvigorating the exhausted T cells in patients. However, the therapeutic efficacy is largely confined due to the primary or acquired resistance to anti-PD-1 (L1) therapy in many patients. Recently, the mechanisms of resistance to ICIs has been extensively elucidated from different aspects and emerging sequencing data from clinical samples has pointed to IFN-γ signaling defects and antigen presentation loss in patients who are resistant to PD-1(L1) blockade. The loss-of-function mutations in JAK1 and JAK2 results in lack of response to IFN-γ signaling and incapacity to upregulate PD-L1 and MHC-I, which subsequently leads to noninflammatory TMEs and resistance to anti-PD-1/PD-L1. The loss-of-function of B2M is likely a common resistance mechanism that the intratumoral cytotoxic CD8+ T cells are failed to be boosted owing to antigen presentation loss. In order to obtain higher clinical benefits in various cancers, it’s appealing and urgent to develop mechanism-based strategies to overcome resistance to ICI-based immunotherapy. To this end, we developed a genetically acquired resistant tumor model with B2M mutations in syngenic MC38 tumor cells using CRISPR/Cas9 technology, which is ‘hot’ tumor to PD-1/PD-L1 blockade. The expression of B2M was detected by western blotting and flow cytometry. Mouse B2M-knockout MC38 tumors became resistant to PD-1/PD-L1 blockade in vivo. Notably, we found a remarkable increase of CD8+ T cell infiltration in B2m-KO MC38 tumors in comparison to parental MC38 tumors upon anti-PD-1 treatment, which further explains that antigen presentation loss results in inactivation of cytotoxic CD8+ T cells within the tumor microenvironments (TMEs), subsequently leading to resistance and poor survival to ICI therapy.In summary, our data consistently indicated the importance of MHC-I expression in T cell activation in the TMEs and the possibility to dig out the pathway involved in MHC-I expression independent of IFN-γ signaling by CRISPR-based screening to overcome MHC-I deficiency-induced resistance in the future. In addition, it’s possible to leverage the cytotoxic NK cells and CD4+ T cells in antitumor immunity against MHC-II+ tumors even with the loss of MHC-I molecules. For example, high dose of IL-2RB-biased IL-2 agonist preferentially binding to the dimeric IL-2R can stimulate Teff and NK cells independent of immune checkpoint expression. Likewise, cytokine modified anti-PD-1 fusion formulation like anti-PD-1-IL-2 mutein and anti-PD-1-IL-15 mutein are promising to overcome the resistance. Furthermore, it has been suggested that the resistance due to genetic mutations of MHC-I could be overcome by combining NF-B targeted therapies. Citation Format: Yanlei Zhang, Hechun Ma, Boang Han, Yi Li, Ping Yang, Zhen Li, Dongxiao Feng, Lei Ci, Ruilin Sun, Daniel X. He. Genetic ablation of B2M leads to resistance to PD-1/PD-L1 blockade in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2666.

New York esophageal squamous cell carcinoma-1 (NY-ESO-1)-specific T cell receptor (TCR) T cell therapy is effective in tumors with NY-ESO-1 expression, but a safe and effective TCR-T cell therapeutic protocol remains to be improved. Here, we report a phase 1 investigational new drug clinical trial with TCR affinity-enhanced specific T cell therapy (TAEST16001) for targeting NY-ESO-1. Enrolled patients receive TAEST16001 cell infusion after dose-reduced lymphodepletion with cyclophosphamide (15 mg/kg/day 3 3 days) combined with fludarabine (20 mg/m2/day 3 3 days), and the TCR-T cells are maintained with low doses of interleukin-2 injection post-adoptive transfer. Analysis of 12 patients treated with the regimen demonstrates no treatment-related serious adverse events. The overall response rate is 41.7%. The median progression-free survival is 7.2 months, and the median duration of response is 13.1 months. The protocol of TAEST16001 cells delivers a safe and highly effective treatment for patients with advanced soft tissue sarcoma (ClinicalTrials.gov: NCT04318964).

BACKGROUND:Although immunotherapy has been considered as a potent strategy for lung adenocarcinoma (LUAD), only a small part of patients was served as potentially clinical benefiters. Immunogenic cell death (ICD), a type of regulated cell death (RCD), which enable to reshape the tumor immune microenvironment and contribute to the immunotherapy efficiency. Developing a novel ICD-based signature may be a potential strategy to differentiate prognosis of patients with LUAD and predict efficacy of immunotherapy.METHODS:In this study, 34 ICD-related genes (ICDRGs) were identified and analyzed in LUAD samples from the Cancer Genome Atlas (TCGA). 572 patients with LUAD were divided into two distinct clusters according to ICDRGs expression levels. Patients were subsequently classified into two distinct gene subtypes based on differentially expressed genes (DEGs) analyzed between two ICD-related clusters. We further developed and validated a novel ICD-related score (ICDRS) followed by comprehensive investigation about the landscape of the prognosis, immune-based features, immunotherapautic responses and sensitivity of target drugs in patients with LUAD.RESULTS:After confirming transcriptomic aberrations and appraising prognostic value of ICDRGs, two ICD-associated subtypes were initially determined by consensus clustering in accordance with differentially expressional levels of ICDRGs. It was shown that patients in the ICD high-subtype possessed the superior clinical prognosis, abundant immune cell infiltration and higher involvement in immune-related signaling compared with the ICD low-subtype. A signature of ICD-related score (ICDRS) was further established and validated, which was served as an independent prognostic indicator for LUAD patients. These comprehensive results revealed that the high-score patients represented better clinical prognosis, higher immune infiltration-related characteristics, stronger expression of immune checkpoints, and better response to immune checkpoint inhibitor therapy and multiple targeted drugs. To further verify our analysis, we selected TLR4 as the representative of ICDRGs and evaluated its expression on the lung normal cells and cancer cells in vitro. Then, relative animal experiments were performed in vivo, with results of that the stimulation of TLR4 suppressed the growth of lung cancer.CONCLUSIONS:In conclusion, our comprehensive analysis of ICDRGs in LUAD demonstrated their function in serving as a biomarker of predicting prognosis and clinical effects of immunotherapy and targeted drugs, which is meaningful to improve our understanding of ICDRGs and brought inspirations about evaluating prognosis and developing effective therapeutic strategies to patients with LUAD.

Fig. S1 shows the SAGE1 expression in NSCLC samples. Fig. S2 shows the PCR products visualized by gel electrophoresis. Fig. S3 shows the proportion of SAGE1597-606-HLA-A24 specific population before and after first stimulation in CD8+ T cells. Fig. S4 shows the FACS staining of non-SAGE1 specific T cells. Fig. S5 shows the KD and Tm values of SAGE1597-606-specific TCRs refolded in vitro. Fig. S6 shows the activation and cytotoxic activity of non-transduced or an irrelevant TCR or GFP-transduced T cells against tumor cells. Fig. S7 shows the number of apoptotic tumor cells in co-cultured with GFP- transduced T cells. Fig. S8 shows the confluence of remaining tumor cells at the endpoint of the IncuCyte assay. Fig. S9 shows the SAGE1 expression in 95D xenograft. Fig. S10 shows the HLA-A11 peptide-specific CTLs induced from two donors. Fig. S11 shows the increased activation and cytotoxic activity of VF3 mutants transduced T cells. Table S1 shows the SAGE1 expression in tumor cells measured by Nanostring nCounter analysis. Table S2 shows the information of three SAGE1597-606-specific TCRs. Table S3 shows the binding of VF TCRs to a panel of irrelevant pHLAs.

Studies have shown that tripartite motif-containing (TRIM) family proteins function as E3 ubiquitin ligases and play essential roles in cancer biology. In the present study, we validated a contribution of TRIM9 to bladder cancer progression. 296 patients derived from The Cancer Genome Atlas (TCGA) database and 22 clinical specimens were included, in which accumulated TRIM9 correlated with the poor prognosis and higher relapse in bladder patients. In vitro, TRIM9 promoted bladder cancer cells Biu-87 and T24 cell proliferation and migration. Meanwhile, overexpression of TRIM9 reduced the chemosensitivity in Biu-87 and T24 to mitomycin C (MMC) and gemcitabine (GEM). As an underlying mechanism, we found that TRIM9 stimulated carcinoembryonic antigen 6 (CEACAM6) upregulation, which further facilitated Smad2/3-matrix metalloproteinase 2 (MMP2) signaling activation both in vitro and in vivo. Those results indicated that TRIM9 facilitated bladder cancer development and chemoresistance by CEACAM6-Smad2/3 axis. TRIM9 and its associated molecules could be a potential diagnostic indicator and therapeutic target in bladder cancer.

IntroductionThe presence of soluble human programmed cell death-ligand 1 (shPD-L1) in the blood of patients with cancer has been reported to be negatively correlated with disease prognosis. However, little information exists about the mechanisms underlying high levels of shPD-L1 for promoting disease progression. MethodsIn this study, we first analyzed the correlations between shPD-L1 and apoptosis of T cells in patients with cancer, then tested the effect of shPD-L1 on T-cell functions and the production of regulatory T cells. ResultsWe found that the apoptosis of human peripheral PD-1+CD4+ T cells was significantly elevated in patients with cancer compared with healthy donors and was positively correlated with circulating PD-L1 levels in patients with cancer. In vitro, monomeric shPD-L1 significantly inhibited the proliferation, cytokine secretion, and cancer cell-killing activity of peripheral blood mononuclear cells (PBMCs) activated by either agonist antibodies or HATac (high-affinity T cell activation core)-NYE (NY-ESO-1 antigen). It also promoted CD4+ T cells to express forkhead family transcription factor 3 (FoxP3) for the conversion of induced T regulatory cells, which was more significant than that mediated by soluble human PD-L1 fusion protein (shPD-L1-Fc). DiscussionThese results confirm that soluble PD-L1 could be a candidate for inhibiting the functions of activated T cells, promoting peripheral tolerance to tumor cells, and implicating in system tumor immune escape in addition to the tumor microenvironment. This is an important mechanism explaining the negative correlation between peripheral blood PD-L1 levels and cancer prognosis. Therefore, understanding the roles of hPD-L1 in peripheral blood will be helpful for the development of precision immunotherapy programs in treating various tumors.

Worldwide, the incidence rate of breast cancer is the highest in women. Approximately 2.3 million people were newly diagnosed and 0.685 million were dead of breast cancer in 2020, which continues to grow. Triple-negative breast cancer (TNBC) is the most aggressive breast cancer subtype with a higher risk of recurrence and metastasis, but disappointly, there are no effective and specific therapies clinically, especially for patients presenting with metastatic diseases. Therefore, it is urgent to develop a new type of cancer therapy for survival improvisation and adverse effects alleviation of breast cancers. Near-infrared photoimmunotherapy (NIR-PIT) is a newly developed, photochemistry-based cancer therapy. It was drive by an antibody–photoabsorber conjugate (APC) which is triggered by near-infrared light. The key part of APC is a cancer-targeting monoclonal antibody (mAb) that can bind to receptors or antigens on the surface of tumor cells. Because of this targeted conjugate accumulation, subsequent deployment of focal NIR-light results in functional damage on the targeted cell membranes without harming the immediately adjacent receptor-negative cells and evokes a kind of photochemical, speedy, and highly specific immunogenic cell death (ICD) of cancer cells with corresponding antigens. Subsequently, immature dendritic cells adjacent to dying cancer cells will become mature, further inducing a host-oriented anti-cancer immune response, complicatedly and comprehensively. Currently, NIR-PIT has progressed into phase 3 clinical trial for recurrent head and neck cancer. And preclinical studies have illustrated strong therapeutic efficacy of NIR-PIT targeting various molecular receptors overexpressed in breast cancer cells, including EGFR, HER2, CD44c, CD206, ICAM-1 and FAP-α. Thereby, NIR-PIT is in early trials, but appears to be a promising breast cancer therapy and moving into the future. Here, we present the specific advantages and discuss the most recent preclinical studies against several transmembrane proteins of NIR-PIT in breast cancers.

Abstract Current clinical breakthroughs in gene therapy have brought adeno-associated virus (AAV) vectors to the forefront of gene delivery systems. Vitamin C deficiency due to GULO mutations is a genetic disorder affecting guinea pigs and humans. In our study, we used AAV9-mGULO-GT to deliver the mouse GULO gene to guinea pigs and restore Vc synthesis in affected tissues, including the liver and brain. AAV9-mGULO-GT treatment significantly improved survival rates and bone health compared to non-treated and Vc-treated groups. Dot blot analysis confirmed restored Vc content in various parts of the brain. Additionally, micro-CT imaging showcased significant enhancements in bone mineral density, content, width, and cortical thickness. Further, RNA sequencing and immunological studies of organs validated the successful restoration of Vc synthesis. These findings highlight the potential of AAV9-mGULO-GT as a therapeutic option for GULO-related scurvy and other genetic disorders. The success of our study underscores the importance of advanced targeting and gene rescue systems in developing effective therapies for genetic disorders in clinical applications.