Biology subjects

Garcia-Donas, J.

Publications and source records attributed to Garcia-Donas, J..

2 recordsLinked to original sources

bioRxiv2025

Identification of a novel papillary renal cell carcinoma molecular subtype characterized by HIF-pathway over-activation

PurposePapillary renal cell carcinoma (pRCC), the second most common subtype of renal cancer, exhibits heterogeneity in molecular features and response to targeted therapies, including antiangiogenic drugs. Discovering molecular biomarkers able to stratify pRCC patients into clinically relevant subgroups and understanding the underlying mechanisms are urgently needed to advance precision medicine. Here, we molecularly dissect a large pRCC series through the expression of the targets of hypoxia inducible factors (HIF), master regulators of angiogenesis, metabolic reprogramming and immune microenvironment. Experimental DesignWe merged and analyzed multi-omic and clinical data of 346 patients derived from two pRCC series (TCGA-KIRP and a Spanish metastatic series). Altered pathways and differences in tumor microenvironment were identified through tumor transcriptomic analyses. Tumor metabolome analysis was performed in selected cases. ResultsMolecular revision of driver mutations classified 302 patients as pRCC, while uncovering misclassified cases. Analysis of HIF targets gene expression identified a subset of pRCC tumors with increased HIF activity (31%; "HIF-active"). These tumors were characterized by high hypoxia scores, increased angiogenesis, low expression of Krebs cycle genes and mitochondrial activity, high immune infiltration and increased epithelial-mesenchymal transition (P<0.0001; each feature) and they were associated with increased metastasis risk and worse overall survival (P<0.005). Metabolome analyses revealed that HIF-active tumors accumulated L-2-hydroxyglutarate (L-2-HG), an oncometabolite that leads to pseudohypoxia by inhibiting HIF prolyl hydroxylases and preventing HIF degradation. L-2-HG-accumulation agreed with diminished L2HGDH expression, associated with losing one copy of the gene and low expression of PPARGC1A, which regulates its transcription. ConclusionsHIF-pathway overactivation defines a pseudohypoxic pRCC molecular subgroup characterized by high angiogenesis, immune infiltration and aggressive features. These findings advance our understanding of pRCC diversity, revealing potential clues for the variable response of patients to targeted therapies and paving the way to more personalized pRCC treatments.

cancer biology↗

bioRxiv2024

CRISPR targeting of FOXL2 c.402C>G mutation reduces malignant phenotype in granulosa tumor cells and identifies anti-tumoral compounds

FOXL2 is a transcription factor essential for sex determination and ovary development and maintenance. Mutations in this gene are implicated in syndromes involving premature ovarian failure and granulosa cell tumors (GCTs). This rare cancer accounts for less than 5% of diagnosed ovarian cancers and is causally associated with the FOXL2 c.402C>G, p.C134W mutation in 97% of the adult cases (AGCTs). In this study, we employed CRISPR technology to specifically eliminate the FOXL2 c.402C>G mutation in granulosa tumor cells. Our results show that this Cas9-mediated strategy selectively targets the mutation without affecting the wild-type allele. Granulosa cells lacking the FOXL2 c.402C>G exhibit a reduced malignant phenotype, with significant changes in cell proliferation, invasion, and cell death. Furthermore, these modified cells are more susceptible to Dasatinib and Ketoconazole. Transcriptomic and proteomic analyses reveal that CRISPR-modified granulosa tumor cells shift their expression profiles towards a wild-type like phenotype. Additionally, this altered expression signature has led to the identification of new compounds with antiproliferative and pro-apoptotic effects on granulosa tumor cells. Our findings demonstrate the potential of CRISPR technology for the specific targeting and elimination of a mutation causing granulosa cell tumors, highlighting its therapeutic promise for treating this rare ovarian cancer. Simple SummaryAdult granulosa cell tumors (AGCTs), characterized by a specific point mutation (C134W) in the gene FOXL2, are less than 5% of all the diagnosed ovarian cancers. Surgery is the cornerstone treatment for AGCT even at relapse, with systemic therapy showing poor results. The aim of our study is to explore the potential therapeutic effect of the elimination of the C134W mutation. To achieve our goal, we have eliminated the mutant allele using CRISPR technology. Our results demonstrate that CRISPR-mediated elimination of FOXL2-C134W reduces the malignant phenotype of granulosa tumor cells, which change their transcriptional, proteomic, and cellular phenotype to a wild type like, granulosa type. Moreover, the induced changes allowed us to find new compounds with antitumoral activity. This work highlights the therapeutic potential of CRISPR mediated technology for the treatment of AGCT.

cancer biology↗