TGFbeta诱导：新一代SERD在体外ER+乳腺癌模型中的一个必要的肿瘤抑制机制臂

Breast cancer is the most diagnosed cancer globally, with a significant need for improved treatments for the most prevalent Estrogen Receptor alpha (ER)-positive subtype, where ER signaling drives cell proliferation.The next-generation oral Selective ER antagonist and degrader (SERD), giredestrant, is a potent investigational agent that robustly suppresses tumor cell proliferation. However, primary and acquired resistance pose a major challenge to all endocrine therapies including oral SERDs, necessitating a deeper understanding of resistance mechanisms. In a comprehensive whole-genome CRISPR screen, we sought to identify genetic perturbations that confer a growth advantage in the presence of giredestrant. Notably, knockout of multiple TGFbeta signaling components (TGFbeta3, SMAD4, IPO8, TGFbetaR2) consistently freed ER+ cells from giredestrant-induced cell cycle arrest. We establish an unexpected autocrine mechanism: by suppressing the ER pathway, giredestrant induces the expression and signaling of the tumor suppressor TGFbeta3. This induced TGFbeta signaling then elevates CDKN1A (p21) levels, which precedes the inhibition of its natural target, CDK2. We demonstrate that this TGFbeta→p21-|CDK2 axis is necessary for giredestrant to achieve its maximal inhibitory effect in vitro. Molecular profiling confirmed CDKN1A was among the most significantly suppressed genes when TGFbeta signaling was inhibited in combination with giredestrant. Crucially, combining giredestrant and a TGFbeta inhibitor led to resistance, but this was partially overcome by a combination of CDK4/6 and CDK2 inhibition. This finding shows that CDK4/6/2 blockade can bypass, in-part, the requirement for the p21 cell cycle brake. Collectively, our results reveal that giredestrant's in vitro efficacy relies on an autocrine TGFbeta loop to mediate the necessary G1 arrest. The inhibition of TGFbeta creates a resistance mechanism by neutralizing this essential tumor-suppressive signaling component.