Molecular characterization of chromophobe renal cell carcinoma reveals mTOR pathway alterations in patients with poor outcome

Roldán-Romero, Juan María; Santos, María; Lanillos, Javier; Caleiras, Eduardo; Anguera, Geòrgia; Maroto, Pablo; García-Donás, Jesús; Velasco, Guillermo; Martínez-Montes, Ángel M; Calsina, Bruna; Monteagudo, María Jesús; Letón, Rocío; Leandro‐García, Luis Javier; Montero‐Conde, Cristina; Cascón, Alberto; Robledo, Mercedes; Rodríguez‐Antona, Cristina

doi:10.1038/s41379-020-0607-z

Cited by 35 publications

(28 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although these differences in GPNMB protein expression among the common RCC subtypes were not statistically significant, the trends in these data parallel the statistically significant differences in GPNMB mRNA expression observed in the TCGA cohorts, where ccRCC cases have the lowest normalized counts, followed by papRCC and chRCC cases (supplementary material, Figure S4) [39–41]. The higher GPNMB expression among at least a subset of chRCCs compared with other common subtypes could potentially be consistent with the previously reported increased frequency of TSC1/TSC2 / MTOR alterations among eosinophilic chRCC cases, ranging from 6% of cases in TCGA to 17% in a metastatic chRCC cohort [41–43]. Within the TCGA chRCC cohort (KICH) [41], a previous study has demonstrated that high FOXI1 mRNA expression is characteristic of chRCC, and outlier cases lacking FOXI1 expression (suggestive of misclassification) are enriched for TSC1/TSC2/MTOR alterations (present in 4/6 cases with low FOXI1 expression) [42].…”

Section: Resultssupporting

confidence: 85%

GPNMB expression identifies TSC1/2/mTOR‐associated and MiT family translocation‐driven renal neoplasms

Salles

Asrani

Woo

et al. 2022

The Journal of Pathology

View full text Add to dashboard Cite

GPNMB (glycoprotein nonmetastatic B) and other TFE3/TFEB transcriptional targets have been proposed as markers for microphthalmia (MiT) translocation renal cell carcinomas (tRCCs). We recently demonstrated that constitutive mTORC1 activation via TSC1/2 loss leads to increased activity of TFE3/TFEB, suggesting that the pathogenesis and molecular markers for tRCCs and TSC1/2-associated tumors may be overlapping. We examined GPNMB expression in human kidney and angiomyolipoma (AML) cell lines with TSC2 and/or TFE3/TFEB loss produced using CRISPR-Cas9 genome editing as well as in a mouse model of Tsc2 inactivation-driven renal tumorigenesis. Using an automated immunohistochemistry (IHC) assay for GPNMB, digital image analysis was employed to quantitatively score expression in clear cell RCC (ccRCC, n = 87), papillary RCC (papRCC, n = 53), chromophobe RCC (chRCC, n = 34), oncocytoma (n = 4), TFE3-or TFEB-driven tRCC (n = 56), eosinophilic solid and cystic RCC (ESC, n = 6), eosinophilic vacuolated tumor (EVT, n = 4), and low-grade oncocytic tumor (LOT, n = 3), as well as AML (n = 29) and perivascular epithelioid cell tumors (PEComas, n = 8). In cell lines, GPNMB was upregulated following TSC2 loss in a MiT/TFE-and mTORC1-dependent fashion. Renal tumors in Tsc2 +/À A/J mice showed upregulation of GPNMB compared with normal kidney. Mean GPNMB expression was significantly higher in tRCC than in ccRCC (p < 0.0001), papRCC (p < 0.0001), and chRCC (p < 0.0001). GPNMB expression in TSC1/2/MTOR alteration-associated renal tumors (including ESC, LOT, AML, and PEComa) was comparable to that in tRCC. The immunophenotype of tRCC and TSC1/2/MTOR alteration-associated renal tumors is highly overlapping, likely due to the increased activity of TFE3/TFEB in both, revealing an important caveat regarding the use of TFE3/TFEB-transcriptional targets as diagnostic markers.

show abstract

Section: Resultssupporting

confidence: 85%

GPNMB expression identifies TSC1/2/mTOR‐associated and MiT family translocation‐driven renal neoplasms

Salles

Asrani

Woo

et al. 2022

The Journal of Pathology

View full text Add to dashboard Cite

show abstract

“…Most recently, Roldan-Romero et al. [ 72 ] found within the eosinophilic variant of ChRCCs an overrepresentation of mTOR pathway ( MTOR , TSC1 , and TSC2 ) mutations as well as aberrations in the mitochondrial genes encoding the complex I of the electron respiratory chain.…”

Section: Molecular Underpinnings Of Chrccsmentioning

confidence: 99%

Chromophobe renal cell carcinoma: Novel molecular insights and clinicopathologic updates

Alaghehbandan

Przybycin

Verkarre

et al. 2022

Asian Journal of Urology

View full text Add to dashboard Cite

“…A detailed description of these three patients is provided in Supplementary Methods. Tumor mutations in genes frequently altered in chRCC and immunohistochemistry characteristics for these three cases and a large series of 89 chRCC patients, was recently published by our group (14).…”

Section: Patientsmentioning

confidence: 84%

“…To study USP9X mutational prevalence in chRCC, we collected 89 additional tumors enriched in metastatic cases (14). All these tumors showed a positive USP9X IHC staining and 3 of 84 cases with mutation data (3.6%) carried a USP9X missense mutation.…”

Section: Usp9x Mutations In Cancermentioning

confidence: 99%

“…A comparison of the clinical characteristics of the USP9Xmutated and wild type patients did not reveal statistically signi cant differences in stage, metastatic status nor survival of the patients. Clinical details, together with the mutational landscape and IHC characteristics of this chRCC series was recently published (14). In the TCGA chRCC series, 2 of 65 tumors (3.0%) were mutated in USP9X with a missense and a frameshift variant (Fig S3).…”

Section: Usp9x Mutations In Cancermentioning

confidence: 99%

See 1 more Smart Citation

Deubiquitinase USP9X loss Sensitizes Cancer Cells to mTOR Inhibitors through p62 Dysregulation

Roldán-Romero

Santos

Lanillos

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

BackgroundMutations in MTOR and TSC1/2 can only explain part of variability in mTOR inhibitor response. Here, we performed a comprehensive molecular characterization of tumors with high sensitivity to these drugs to uncover novel mechanisms of response.MethodsChromophobe renal cell carcinoma (chRCC) tumors were analyzed by whole-exome sequencing. Rapamycin sensitivity, expression of mTOR pathway effectors, ubiquitylome analyses to identify USP9X substrates, p62 immunoprecipitation and autophagy assessment through immunofluorescence assays for p62 and LC3 were performed in USP9X depleted HeLa and 786-O cells for mechanistic investigation. ResultsWhole-exome sequencing of chRCC patients with high mTOR inhibitor sensitivity, uncovered USP9X deubiquitinase as the only mutated gene shared by these tumors. The clonal characteristics of the mutations, revealed by studying multiple primary and metastatic samples, together with the low USP9X mutation rate in unselected chRCC series, implied a causal link between USP9X and mTOR inhibitor sensitivity. The high sensitivity was recapitulated in vitro, and while no direct effect on mTORC1 was detected, an unbiased ubiquitylome analysis revealed p62 as a direct USP9X target. Bortezomib treatment, which also led to p62 ubiquitination, increased rapamycin effect. Finally, immunofluorescence assays for p62 and LC3 confirmed dysregulated autophagy in USP9X depleted cells, supporting a synthetic lethal interaction between rapamycin-induced autophagy via mTOR-axis and USP9X loss.ConclusionsOur study uncovers USP9X as a potential novel marker of sensitivity to mTOR inhibitors and suggests the inhibition of this gene as a clinically exploitable strategy able to increase sensitivity to these drugs through a novel p62 regulatory mechanism.

show abstract

Molecular characterization of chromophobe renal cell carcinoma reveals mTOR pathway alterations in patients with poor outcome

Cited by 35 publications

References 33 publications

GPNMB expression identifies TSC1/2/mTOR‐associated and MiT family translocation‐driven renal neoplasms

GPNMB expression identifies TSC1/2/mTOR‐associated and MiT family translocation‐driven renal neoplasms

Chromophobe renal cell carcinoma: Novel molecular insights and clinicopathologic updates

Deubiquitinase USP9X loss Sensitizes Cancer Cells to mTOR Inhibitors through p62 Dysregulation

Contact Info

Product

Resources

About