Aug 2026· Cancer Science· 0 citations· 40 references
Medicine
TL;DR
The T-Editor platform enables rapid and efficient CRISPR-mediated gene editing for engineering TIL to enhance its therapeutic potency and FAM84B may represent a novel potential target for improving TIL-mediated antitumor activity.
Abstract
ABSTRACT Tumor‐infiltrating lymphocytes (TIL) therapy has demonstrated clinical potential in malignancies. However, limited understanding of why only a subset of patients respond to TIL therapy, coupled with the lack of simple and efficient methods to genetically engineer fragile TIL, has hindered efforts to enhance TIL efficacy through genetic modification. A T‐Editor platform enabling rapid and efficient CRISPR‐mediated gene editing in TIL was developed and optimized. To minimize the risk of chromosomal translocations associated with Cas9‐induced double‐strand breaks (DSBs), single‐guide RNAs (sgRNAs) were designed for cytosine base editing (CBE). The expansion capacity, phenotypic profile, cytokine production, and in vitro cytolytic activity of base‐edited TIL were compared with those of Cas9‐KO TIL. In vivo efficacy was assessed using patient‐derived xenograft (PDX) mouse models. The T‐Editor platform was optimized for TIL gene editing by refining stimulation conditions, electroporation parameters, and CRISPR/Cas9 reagent dosing. FAM84B emerged as the top candidate, with its knockout resulting in the most pronounced enhancement of TIL cytolytic activity. CBE‐mediated C·G‐to‐T·A conversion in the FAM84B exon achieved high editing efficiency with minimal insertion–deletion (indel) events. Base‐edited TIL exhibited comparable expansion, phenotype, cytokine production, and in vitro cytolytic activity relative to Cas9‐KO TIL. Compared with non‐engineered control TIL, FAM84B‐edited TIL displayed an increased CD62L+ memory subset, enhanced effector function and cytolytic activity, and improved in vivo antitumor efficacy. In conclusion, the T‐Editor platform enables rapid and efficient CRISPR‐mediated gene editing for engineering TIL to enhance its therapeutic potency. FAM84B may represent a novel potential target for improving TIL‐mediated antitumor activity.
ABSTRACT Chimeric antigen receptor (CAR) T‐cell therapy has achieved durable efficacy in hematologic malignancies but encounters persistent obstacles in solid tumours, including antigen heterogeneity, a suppressive tumour microenvironment (TME), and intrinsic T‐cell dysfunction. This review examines the transition from single‐axis engineering to an integrated framework that addresses these hurdles in sequence. We delineate how next‐generation CAR‐T cells are designed for precise spatiotemporal activation through logic‐gated and pharmacologically regulatable receptors, while being reinforced by metabolic and epigenetic reprogramming to resist TME‐driven exhaustion. We also assess strategies that actively reshape the immunosuppressive TME, including depletion of regulatory cell populations, blockade of ‘don't eat me’ signals, and the use of biomaterial scaffolds for locoregional delivery. The synthesis of controllable activation, intrinsic resilience, and extrinsic TME modulation is defining a class of adaptive therapeutic systems. Clinical implementation of this approach requires careful management of toxicities, notably cytokine release syndrome (CRS), and support from advanced monitoring technologies. Progress will depend on rational combinations that move beyond isolated optimisations, enabling cellular therapies to dynamically respond to evolving tumour ecosystems and narrowing the efficacy gap between hematologic and solid cancers.
Chao Yang, Tan Li, Ping He et al.· Cell Proliferation· 0 citations
Immuno-oncology has reshaped the therapeutic landscape of cancer treatment by shifting focus from directly targeting tumor cells to mobilizing the immune system against malignancies. Among the most transformative advances in this field is the development of chimeric antigen receptor T-cell therapy, which has demonstrated remarkable efficacy in hematologic cancers. However, persistent challenges such as limited durability, immune escape, toxicity, and poor performance in solid tumors have constrained its broader clinical impact. The emergence of clustered regularly interspaced short palindromic repeats (CRISPR) genome editing has introduced a powerful and versatile platform for engineering immune cells with enhanced specificity, persistence, and functionality. CRISPR-based approaches enable precise gene knockout, targeted gene insertion, epigenetic modulation, and multiplex editing, allowing researchers to redesign immune cells at multiple regulatory levels. These capabilities have significantly advanced CAR-T cell engineering and have catalyzed the development of next-generation immune effectors, including natural killer cells, macrophages, and stem cell-derived immune populations. Furthermore, CRISPR technology has opened new avenues for overcoming the immunosuppressive tumor microenvironment, improving safety profiles, and enabling scalable, off-the-shelf therapies. This review provides a comprehensive examination of CRISPR applications in immuno-oncology, with an emphasis on CAR-T optimization and the engineering of next-generation immune cells. It discusses mechanistic foundations, technological innovations, preclinical and clinical advancements, safety considerations, and future directions. Collectively, CRISPR-driven immune engineering represents a paradigm shift toward more precise, effective, and accessible cancer immunotherapies.
Adewale Adeleke· International Journal for Sc...· 0 citations
Although chimeric antigen receptor T (CAR-T) cell therapy has achieved remarkable success in hematological malignancies, its therapeutic efficacy in solid tumors remains limited by several challenges, including insufficient tumor infiltration, T cell exhaustion and the immunosuppressive tumor microenvironment (TME). CRISPR/Cas, a third-generation gene editing technology developed in recent years, is characterized by its simplicity and high efficiency. This technology has demonstrated broad application potential across multiple fields and has emerged as a powerful tool for improving CAR-T cell therapy. In this review, we summarize recent advances in the application of CRISPR/Cas gene editing technology to enhance the antitumor activity of CAR-T cells against solid tumors. We also discuss the key challenges currently faced and systematically propose potential strategies for overcoming the limitations.
Wenjing Liu, Jiayi Gu, Chenghao Xie et al.· Frontiers in Immunology· 0 citations
The advent of CRISPR/Cas9 genome editing has significantly transformed the landscape of cancer therapeutics by facilitating precise and programmable manipulation of disease-associated genetic modifications. This review comprehensively evaluates the current clinical and translational landscape of CRISPR/Cas9-based cancer therapies through an analysis of published literature and registered clinical trials. The current CRISPR/Cas9 applications in oncology are primarily centred on three mechanistic strategies: immune cell engineering for enhanced tumor recognition, direct targeting of oncogenic mutations, and modulation of tumor-supportive pathways. Analysis of 32 clinical trials indicates that CRISPR-based interventions have demonstrated encouraging safety profiles and early signs of clinical activity, particularly in ex vivo engineered immune-cell therapies. Notable examples include CRISPR-edited CAR-T cell products targeting CD19 and BCMA, which have achieved objective responses in relapsed or refractory hematological malignancies while demonstrating sustained persistence of edited cells in vivo. In contrast, clinical translation into solid tumors remains comparatively limited due to challenges associated with delivery efficiency, tumor heterogeneity, and the immunosuppressive tumor microenvironment. Technological advancements, including multiplex genome editing, base editing, and prime editing have expanded the precision and versatility of CRISPR-based interventions, while integration with immunotherapy and nanotechnology-based delivery systems continues to broaden therapeutic potential. Despite these advances, several significant challenges still need to be addressed, including off-target editing, manufacturing scalability, delivery limitations, and regulatory considerations. Overall, CRISPR/Cas9 represents a promising yet evolving platform in oncology, with its future clinical success dependent on achieving a balance between precision, safety, scalability, and long-term therapeutic durability.
Chu Xin Ng, Sakina Mustafa, X. Y. Yap et al.· Frontiers in Oncology· 0 citations
Abstract Objective Anti‐PD‐1 therapy resistance remains a critical barrier in non‐small cell lung cancer (NSCLC) management, and the underlying mechanisms are incompletely defined. Methods We generated CD155‑knockout (KO) NSCLC cell lines using the CRISPR‑Cas9 system and performed systematic multi‑omics analyses, including single‑cell RNA‑seq, bulk RNA‑seq, proteomics, and metabolomics. The key molecular mechanisms were further validated by immunohistochemistry (IHC), western blotting, and chromatin immunoprecipitation (ChIP). Functional assays assessed cell proliferation, migration, and metabolic phenotypes, while the therapeutic efficacy was assessed in vivo using AAV9_shCD155. Results Single‐cell sequencing revealed aberrantly high CD155 expression in NSCLC patients with poor response to anti‐PD‐1 therapy. High CD155 expression in NSCLC tissues correlated with unfavourable prognosis. ETS1 was identified as a direct transcriptional driver of CD155. Multi‐omics analysis and functional assays demonstrated that CD155 upregulates the expression of key glycolytic proteins (GLUT1, GLUT3, LDHB) by activating the PI3K/AKT/HIF‐1α signalling axis, thereby driving glycolytic metabolism, proliferation, and migration of tumour cells. CD155 knockout significantly suppressed these malignant phenotypes. In xenograft mouse models, monotherapy with AAV9_shCD155 effectively inhibited tumour growth and postoperative recurrence. More importantly, in humanised mouse models, combining AAV9_shCD155 with pembrolizumab produced synergistic anti‐tumour effects, more significantly suppressing tumour growth and promoting immune cell infiltration into the tumour microenvironment. Conclusion CD155 mediates anti‐PD‐1 resistance by activating PI3K/AKT/HIF‐1α‐driven glycolytic reprogramming. Targeting CD155 combined with anti‐PD‐1 overcomes resistance, supporting a dual‐target therapeutic strategy. Key points CD155 is identified as a key driver of anti‐PD‐1 resistance in NSCLC. CD155 promotes tumour glycolysis and malignant progression via the PI3K/AKT/HIF‐1α signalling axis. AAV9_shCD155 combined with anti‐PD‐1 markedly inhibits tumour growth and promotes immune infiltration.
Weiguang Du, Xi-Yang Tang, Yulong Zhou et al.· Clinical and Translational M...· 0 citations
ABSTRACT Therapy resistance in prostate cancer arises from coordinated remodeling of malignant and stromal compartments, yet the mechanisms orchestrating this ecosystem adaptation remain elusive. Here, single‐cell RNA sequencing of longitudinal biopsies obtained before and after androgen‐deprivation therapy (ADT) delineated a therapy‐induced stromal lineage bifurcation toward APOD+ and DPT+ fibroblast states. DPT+ fibroblasts activated a C3‐ITGAX/ITGB2 complement signaling axis targeting macrophages, coinciding with suppression of M1 inflammatory programs, amplification of immune‐checkpoint signaling, and a shift of CD8+ T cells from cytotoxic to exhausted phenotypes. Concomitantly, we identified pre‐existing malignant epithelial subpopulations characterized by reduced AR/KLK3 activity and heightened chromosomal instability that preferentially persisted following therapy. Integrative multi‐omic analyses nominated TSPAN1 as a functional effector of castrate resistant prostate cancer (CRPC) and NRXN1 as a regulator of neuroendocrine plasticity through calcium‐dependent signaling programs. Genetic silencing of either gene suppressed proliferation, clonogenicity, migration, and tumor growth, while attenuating neuroendocrine features in vitro and in vivo. Spatial mapping, functional perturbation, and stromal‐epithelial co‐culture experiments mechanistically established a therapy‐induced DPT+ fibroblast‐complement circuit that enforced immune evasion and channels epithelial trajectories toward CRPC or neuroendocrine prostate cancer. Collectively, these findings defined the DPT+‐complement‐macrophage axis as an actionable vulnerability and position TSPAN1 and NRXN1 as therapeutic entry points to disrupt ADT‐driven tumor ecosystem remodeling in prostate cancer.
Yang Chen, Dandan Dong, Jinling Liao et al.· Advancement of science· 0 citations
A new method for surgically removing training examples from a model reveals that as datasets grow, the link between what a model learns and what it produces dissolves.
