CRISPR targeting of FOXL2 c.402C>G mutation reduces malignant phenotype
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1132209 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR29714245_1.fastq.gz | Fastqsanger.gz | |||
| SRR29714245_2.fastq.gz | Fastqsanger.gz | |||
| SRR29714246_1.fastq.gz | Fastqsanger.gz | |||
| SRR29714246_2.fastq.gz | Fastqsanger.gz | |||
| SRR29714247_1.fastq.gz | Fastqsanger.gz |
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Project description: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.
2025-04-16 | GSE271552 | GEO
Project description:FOXL2 is a transcription factor that plays a key role in sex determination, ovary development and maintenance. Mutations related to this gene have been described in syndromes involving premature ovarian failure and granulosa cell tumors. This kind of rare cancer (less than 5% of diagnosed ovarian cancers) has been causally associated with the FOXL2 c.402C>G, p.C134W mutation in 97% of the adult cases (AGCTs). In this study, we have used CRISPR technology to specifically eliminate the FOXL2 c.402C>G mutation in granulosa tumor cells. Our results indicate that this Cas9-mediated strategy allows the specific elimination of the mutation with no activity on the wild type allele. Granulosa cells depleted on FOXL2 c.402C>G show a reduced malignant phenotype. Specifically, we detect changes in cell proliferation, invasion, and cell death levels. In addition, we show that granulosa tumor cells become more susceptible to Dasatinib and Ketoconazole treatments when FOXL2 c.402C>G allele is eliminated. Our transcriptomic and proteomic analyses indicate that CRISPR-modified granulosa tumor cells significantly change their expression signature towards a wild type like phenotype. Finally, this expression signature has led us to discover new compounds with antiproliferative and proapoptotic effects on granulosa cell tumor cells. Our results demonstrate the potential of CRISPR for specifically targeting and eliminating a granulosa cell tumor-causing mutation, as well as its therapeutic potential for the treatment of this rare ovarian cancer.
2025-03-15 | PXD053682 | Pride
Project description:The somatic missense point mutation c.402C>G (p.C134W) in the FOXL2 transcription factor is pathognomonic for adult-type granulosa cell tumours (AGCT) and a diagnostic marker for this tumour type. However, the molecular consequences of this mutation and its contribution to the mechanisms of AGCT pathogenesis remain unclear. To explore the mechanisms driving FOXL2C134W pathogenicity we engineered V5-FOXL2 and V5-FOXL2C134W doxycycline inducible isogenic cell lines and performed ChIP-seq and transcriptome profiling following transgenic induction. Interestingly we found that FOXL2C134W associates with the majority of FOXL2 binding sites as well as a large collection of novel regions throughout the genome. De novo motif analysis of the DNA elements uniquely bound by FOXL2C134W revealed a novel DNA motif that was not seen in the shared or FOXL2 specific DNA elements and we validated an altered DNA binding specificity for FOXL2C134W using in vitro band shift assays. Integrated analysis of FOXL2C134W specific proximal promoter binding sites and differential gene expression identified direct targets of FOXL2C134W. We validated one such target and demonstrated a FOXL2C134W dependent sensitivity to a therapeutic that targets survivin (YM155). Taken together our results suggest that FOXL2C134W pathogenicity in AGCT is driven in part by an alteration in DNA binding specificity contributing to an oncogenic transcriptional program.
2020-07-15 | GSE126171 | GEO
Project description:The Foxl2 transcription factor is required for ovarian function during follicular development. Our approach to begin to understand Foxl2 function is through the identification of Foxl2 regulated genes in the ovary. Transiently transfected KK1 mouse granulosa cells were used to identify genes that are potentially regulated by Foxl2. KK1 cells were transfected in three groups (mock, activated, and repressed) and twenty-four hours later RNA was isolated and submitted for Affymetrix microarray analysis. Experiment Overall Design: To increase the potential of Foxl2 to alter gene expression levels of putative target genes, two fusions were constructed consisting of Foxl2 fused to the activation domain of the Herpes simplex virus VP16 transcription factor (Foxl2-VP16) and Foxl2 fused to the repression domain of the murine MAD transcription factor (Foxl2-MAD). Levels of gene expression were compared between mock transfected cells and those that were transfected with Foxl2-VP16 and Foxl2-MAD, respectively.
2009-11-15 | E-GEOD-18891 | biostudies-arrayexpress
Project description:We have generated a humanized Foxl2 mouse model by genome editing to evaluate in vivo the role of mutated FOXL2 in the development of murine Adult Granulosa Cell Tumors
2023-01-31 | GSE202242 | GEO
Project description:The Foxl2 transcription factor is required for ovarian function during follicular development. Our approach to begin to understand Foxl2 function is through the identification of Foxl2 regulated genes in the ovary. Transiently transfected KK1 mouse granulosa cells were used to identify genes that are potentially regulated by Foxl2. KK1 cells were transfected in three groups (mock, activated, and repressed) and twenty-four hours later RNA was isolated and submitted for Affymetrix microarray analysis.
2009-11-05 | GSE18891 | GEO
Project description:FOXL2 is a lineage determining transcription factor in the ovary, but its direct targets and modes of action are not fully characterized. Here, we explore the genomic targets of FOXL2. We found in particular that FOXL2 directly modulates Esr2 expression through a newly identified intronic element. Input DNA and FOXL2 ChIP
2014-10-29 | E-GEOD-60858 | biostudies-arrayexpress
Project description:The identity of the gonads is determined by which fate, ovarian granulosa cell or testicular Sertoli cell, the bipotential somatic cell precursors choose to follow. In most vertebrates, the fate of granulosa cells is controlled by a conserved regulator FOXL2. To understand how FOXL2 elicits its fate-determining action, we performed genome-wide analysis of FOXL2 chromatin occupancy in fetal ovaries. Combining genome-wide analysis of FOXL2 binding in the fetal ovary with transcriptomic analyses of Foxl2 gain-of-function and Foxl2 loss-of-function models, we identified potential pathways responsible for the feminizing action of FOXL2. Finally, comparison of FOXL2 genome-wide occupancy in the fetal ovary with testis-determining factor SOX9 genome-wide occupancy in the fetal testis revealed extensive overlaps, implying that antagonistic signals between FOXL2 and SOX9 occur at the chromatin level.
2019-03-21 | GSE110093 | GEO
Project description:This SuperSeries is composed of the following subset Series: GSE12905: Foxl2 functions in sex determination and histogenesis throughout mouse ovary development, analyzed by Affymetrix arrays GSE12942: Foxl2 functions in sex determination and histogenesis throughout mouse ovary development, analyzed by Agilent arrays Refer to individual Series
2010-05-17 | E-GEOD-12989 | biostudies-arrayexpress
Project description:In order to identify novel FOXL2 targets, we have transfected KGN cells using an expression vector containing the coding sequence of FOXL2 or, as a reference, the empty vector (mock transfection). The transcriptome perturbation induced by FOXL2 overexpression was then followed by DNA chips (i.e. FOXL2- vs. mock-transfected cells) using the platform developed by NimbleGen Systems, Inc.
2007-03-02 | E-MEXP-985 | biostudies-arrayexpress