Biology subjects

Grana, G.

Publications and source records attributed to Grana, G..

3 recordsLinked to original sources

bioRxiv2026

TIM3+ Tumor Associated M2 Macrophages Impair Antitumor T Cell Immunity and Promote Gastric Cancer Progression and Peritoneal Metastasis

Peritoneal metastases (PM) are the leading cause of cancer-related death in gastric cancer (GC) patients with survival typically < 9 months. Here, we demonstrate that TIM3 and its ligands are increased along the GC continuum and associated with poor survival. Integrated omics analyses and functional studies revealed highly enriched TIM3 in CD163+ tumor associated M2 immunosuppressive macrophages significantly promote tumor cell invasion and tumor growth in vivo, while TIM3 depletion in macrophages reduced tumor cell malignant attributes and increased T cell immunity from PBMCs or CD45+ immune cells of malignant ascites in co-culture system. By cytokine and kinase arrays, we discovered that depletion of TIM3 in macrophages reduced the production of notable secretome of cytokines/chemokines from M2 macrophages; and the protumor function of TIM3+ macrophages rely on the p90RSK1/2/CCL20 axis. Finally, we reveal that TIM3 blockage or genetic KO had superior antitumor activity in combination with anti-PD1 immunotherapy and mitomycin C (MMC) chemotherapy. Together, this study uncovers an important role for TIM3 in tumor associated M2 macrophages and underscores the potential of TIM3 blockage in GC patients with PM. Statement of significanceIn this study, we show TIM3 increases along GC continuum, and highly enriched on tumor associated M2 macrophages that fuel tumor growth; and suppress T cell function via p90RSK1/2/CCL20 axis. TIM3 depletion restores T-cell immunity and curbs tumor growth. TIM3 blockade combined with anti-PD1 and mitomycin C provide a novel therapeutic strategy for GC patients with PM.

immunology↗

bioRxiv2025

YAP1 Depletion Enhances TAZ and its Complexation with TEAD4 and AP-1 Heterodimer C-JUN/FOSB to Promote Gastric Cancer Progression and Metastases

BackgroundDysregulation of the Hippo signaling pathway, characterized by aberrant activation of the transcriptional coactivators YAP1 and TAZ, drives tumour progression, immunosuppression and metastasis. Hippo pathway components are emerging therapeutic targets in several solid tumours, however, the expression profiles of Hippo coactivators YAP1, TAZ and their transcriptional factors TEAD1-4 in gastric cancer peritoneal metastases (GCPMs) and their therapeutic value are unknown. ObjectiveTo determine expression status of YAP1, TAZ and TEAD1-4 in GCPMs; and to evaluate whether dual targeting of YAP1 and TAZ provides superior antitumour activity compared with inhibition of either coactivator alone. DesignExpression of YAP1, TAZ and TEAD1-4 was examined in GCPMs by single-cell RNA sequencing and co-immunofluorescent staining. Functional studies using genetic knockout and antisense oligonucleotide (ASO) inhibition of YAP1 or TAZ were performed to assess antineoplastic effects in vitro and in vivo. Co-immunoprecipitation and luciferase reporter assays were used to characterize YAP1/TAZ interactions with TEADs and AP-1 components (JUN and FOSB) and to quantify transcriptional activity. Antitumour efficacy was validated in patient-derived xenograft (PDX) and KP-Luc2 syngeneic models. ResultsYAP1, TAZ, and TEADs1- 4 were highly coexpressed in GCPMs and correlated with poor survival. YAP1 inhibition alone elicited compensatory upregulation of TAZ, while combined inhibition of both coactivators maximally repressed cell proliferation and invasion in vitro, and tumor growth in vivo. Increased TAZ complexation with TEAD4 and AP-1 (c-JUN and FOSB) heterodimer was observed following YAP1 knockdown or pharmacological ASO inhibition. Dual inhibition of YAP1 and TAZ was required to maximally suppress YAP1/TAZ expression and reduce their nuclear accumulation, transactivation of TEAD, and activation of downstream genes. ConclusionsThese findings show that combined YAP1 and TAZ inhibition holds promise for the treatment of GCPMs, a highly lethal disease with an urgent need for novel treatment options. WHAT IS ALREADY KNOWN ON THIS TOPICO_LIGastric cancer with peritoneal metastasis (GCPM), occurring in > 45% of gastric cancer (GC) patients, is a highly lethal malignancy with limited therapeutic options. C_LIO_LIThe Hippo pathway mediator Yes-associated protein 1 (YAP1) is intimately involved in chemoresistance, cancer stemness properties, and the epithelial-to-mesenchymal transition in gastric and other cancers. C_LIO_LIWhile YAP1 represents a promising therapeutic target, clinical trial of YAP1 antisense oligonucleotides has been disappointing. C_LI WHAT THIS STUDY ADDSO_LIHippo coactivators YAP1, TAZ and their main transcription factors TEAD1-4 are markedly upregulated in GCPMs and associated with poor prognosis. C_LIO_LIAntisense targeting of YAP1 results in compensatory upregulation of its paralog TAZ. C_LIO_LIUpon YAP1 depletion, TAZ forms transcriptional complexes with TEAD4 and the AP-1 heterodimer (JUN and FOSB). C_LIO_LIDual targeting of YAP1 and TAZ, by oligonucleotides, achieves maximal suppression of tumour growth in vitro and in vivo. C_LI HOW THIS STUDY MIGHT AFFECT RESEARCH, PRACTICE OR POLICYO_LIThese findings provide a strong mechanistic rationale for co-targeting YAP1 and TAZ as a therapeutic approach in metastatic gastric cancer. C_LIO_LIDual Hippo coactivator inhibition could inform the design of future clinical trials aimed at improving outcomes for patients with GCPMs. C_LI

cancer biology↗

bioRxiv2025

CDK9 degradation Inhibits Gastroesophageal Cancer growth and Overcomes Radiation Resistance by Increasing Chromatin Accessibility and Downregulating YAP1/TEAD signaling

BackgroundGastroesophageal cancer (GEAC) remains a major health burden and urgently needs novel therapeutic targets. The inhibition of CDK9s activity holds the potential to be a highly effective anti-cancer therapeutic. However, the functional role of CDK9, and its potential targeting in GEAC, remain largely unknown. ObjectiveWe aim to evaluate the potential of degradation CDK9 in GEAC treatment and explore its mechanisms. DesignWe evaluated the expression and distribution of CDK9 in GEAC tissue. We designed and synthesized novel CDK9 degraders using proteolysis targeting chimeras (PROTACs) strategy and selected the promising one for further anti-tumor activity evaluation both in vitro and in vivo. We evaluated the effects of CDK9 degradation on epigenetic reactivation, gene expression and chromatin accessibility. We evaluated the co-targeting of CDK9 and YAP/Tead signaling for GEAC treatment, especially for radiation-resistant tumor treatment. ResultsWe demonstrated significantly elevated CDK9 expression in primary GEAC tumor tissues compared to normal tissues, in association with poor survival. We developed a novel CDK9 degrader, YX0597, reducing RNA Pol II Serine 2 phosphorylation, and inhibition MCL-1; this was accompanied by potent inhibition of GEAC cell growth, especially in radiation-resistant tumor cells. Mechanistically, YX0597 strongly enhanced chromatin accessibility, activating epigenetically silenced genes; and dramatically inhibited YAP/TEAD signaling. CDK9 closely interacts with YAP/TEAD signaling, and co-targeting these two mediators could be a novel treatment strategy for the treatment of GEAC. ConclusionOur studies reveal a new avenue for targeting CDK9-hyperactivated GEAC tumors, especially in combination with YAP1/TEAD inhibition in radiation-resistant GEAC tumors. What is already known on this subject?Gastroesophageal cancer (GEAC), the third-leading cause of global cancer death and urgently needs novel targeted therapies. Targeted protein degradation has emerged as an attractive strategy to fight cancer, complementing the activity of traditional small-molecule inhibitors. Cyclin-dependent kinase 9 (CDK9) has been implicated in various cancers. Recently, CDK9 degraders have now been developed for future clinical benefit in targeting tumors with highly activated CDK9. What are the new findings?In this study, we show that CDK9 expression was significantly elevated in primary GEAC tumor tissues including PDXs compared to normal tissues, in association with poor survival. We developed a novel CDK9 degrader XY0597 and proved that it inhibits the progression of GEAC by increasing chromatin accessibility and down-regulating YAP/Tead signaling. We also proved a strong connection between CDK9 and YAP/TEAD signaling and Co-targeting them synergistically inhibits GEAC tumor cell growth, especially in tumors with high YAP signaling activity, such as radiation-resistant cancer. How might it impact on clinical practice in the foreseeable future?Our data suggests that CDK9 is a viable anticancer target in GEAC. We demonstrate that a novel CDK9 degrader, YX0597, has high potential clinical application for GEAC treatment, especially in radiation-resistant cancer. We reveal the crosstalk of CDK9 and YAP/TEAD signaling, and that co-targeting these two mediators could be a new avenue for targeting CDK9-hyperactivated GEAC primary and radiation-resistant tumors.

cancer biology↗