HIF-2[alpha] expression and metabolic signaling require ACSS2 in clear cell renal cell carcinoma

Clear cell renal cell carcinoma (ccRCC) is an aggressive cancer driven by VHL loss and aberrant HIF-2[alpha] signaling. Identifying means to regulate HIF-2[alpha] thus has potential therapeutic benefit. Acetyl- CoA synthetase 2 (ACSS2) converts acetate to acetyl-CoA and is associated with poor patient prognosis in ccRCC. Here we tested the effects of ACSS2 on HIF-2[alpha] and cancer cell metabolism and growth in ccRCC models and clinical samples. ACSS2 inhibition reduced HIF-2[alpha] levels and suppressed ccRCC cell line growth in vitro, in vivo, and in cultures of primary ccRCC patient tumors. This treatment reduced glycolytic signaling, cholesterol metabolism, and mitochondrial integrity, all of which are consistent with loss of HIF-2[alpha]. Mechanistically, ACSS2 inhibition decreased chromatin accessibility and HIF- 2[alpha] expression and stability. While HIF-2[alpha] protein levels are widely regulated through pVHL-dependent proteolytic degradation, we identify a potential pVHLindependent pathway of degradation via the E3 ligase MUL1. We show that MUL1 can directly interact with HIF-2[alpha] and that overexpression of MUL1 decreased HIF-2[alpha] levels in a manner partially dependent on ACSS2. These findings identify multiple mechanisms to regulate HIF-2[alpha] stability and ACSS2 inhibition as a strategy to complement HIF-2[alpha]-targeted therapies and deplete pathogenically stabilized HIF-2[alpha].