In vivo CRISPR screens identify RhoV as a pro‐metastasis factor of triple‐negative breast cancer

Abstract

Metastasis is the main death reason for triple-negative breast cancer (TNBC). Thus, identifying the driver genes associated with metastasis of TNBC is urgently needed.CRISPR screens have dramatically enhanced genome editing and made it possible to identify genes associated with metastasis. In this study, we identified and explored the crucial role of ras homolog family member V (RhoV) in TNBC metastasis. Here, we performed customized in vivo CRISPR screens targeting metastasis-related genes obtained from transcriptome analysis of TNBC. The regulatory role of RhoV in TNBC was validated using gain-or loss-of-function studies in vitro and in vivo. We further conducted immunoprecipitation and LC-MS/MS to explore the metastasis mechanism of RhoV. In vivo functional screens identified RhoV as a candidate regulator involved in tumor metastasis. RhoV was frequently upregulated in TNBC and correlated with poor survival. Knockdown of RhoV significantly suppressed cell invasion, migration, and metastasis both in vitro and in vivo. In addition, we provided evidence that p-EGFR interacted with RhoV to activate the downstream signal pathway of RhoV, thereby promoting tumor metastasis. We further confirmed that this association was dependent on GRB2 through a specific proline-rich motif in the N-terminus of RhoV. This mechanism of RhoV is unique, as other Rho family proteins lack the proline-rich motif in the N-terminus.

Engineering CRISPR/Cas9 therapeutics for cancer precision medicine

“…In cancer cells (LoVo and SW480), knockout of Aldolase B using CRISPR demonstrated that Aldolase B inhibits proliferation, migration, and invasion in these cancer cells ( Li et al, 2017 ). RhoV is a key driver gene associated with and is upregulated in triple-negative breast cancer ( Jin et al, 2023 ). Studies have shown in vivo functional screens identified RhoV as a regulator of tumor metastasis ( Jin et al, 2023 ).…”

“…RhoV is a key driver gene associated with and is upregulated in triple-negative breast cancer ( Jin et al, 2023 ). Studies have shown in vivo functional screens identified RhoV as a regulator of tumor metastasis ( Jin et al, 2023 ). Jin et al have demonstrated that the knockout of RhoV suppressed cell invasion, migration, and metastasis ( Jin et al, 2023 ).…”

“…Studies have shown in vivo functional screens identified RhoV as a regulator of tumor metastasis ( Jin et al, 2023 ). Jin et al have demonstrated that the knockout of RhoV suppressed cell invasion, migration, and metastasis ( Jin et al, 2023 ). Additionally, it provides evidence that RhoV interacts with p-EGFR to activate the downstream signaling ( Jin et al, 2023 ).…”

“…Jin et al have demonstrated that the knockout of RhoV suppressed cell invasion, migration, and metastasis ( Jin et al, 2023 ). Additionally, it provides evidence that RhoV interacts with p-EGFR to activate the downstream signaling ( Jin et al, 2023 ). Cell division cycle 7 (CDC7) was identified by Deng.…”

Atypical RhoUV GTPases in development and disease

“…RhoUV proteins have also been linked to growth factor receptor tyrosine kinase signaling, primarily through the N-terminal SH3-binding domain. In breast cancer cells, RhoV was shown to directly bind to Grb2, an SH3 domain-containing adapter protein that functions downstream of the epidermal growth factor receptor (EGFR) [ 34 ]. Disrupting the binding between RhoV and Grb2 inhibited EGF-dependent migration.…”