Investigating the role of Cdkn2a in lung adenocarcinoma metastasis

Lung cancer is the most frequent cause of cancer-related deaths in the world. Metastasis is responsible for the overwhelming majority of these deaths, yet the mechanisms underlying metastasis remain poorly understood. This is due in large part to the great difficulty in modeling metastasis for high-throughput functional analyses, as well as recapitulating complex tumor genotypes observed in humans. Using CRISPR-Cas9 somatic genome editing and tumor barcoding, I have developed a novel metastasis assay in genetically engineered mice that can be used to explore functional genetic effects on metastasis and immune evasion. With this approach, I found that inactivation of Cdkn2a promotes metastatic colonization and growth of oncogenic KRAS-driven lung adenocarcinoma. Furthermore, the simultaneous inactivation of several genes along with Cdkn2a dramatically increases metastatic spread, allowing spontaneous metastasis to the liver, bone and brain. In this proposed study, I hypothesize that specific tumor genotypes involving inactivation of Cdkn2a can enable potent metastatic ability. I will explore the ability of Cdkn2a knockout to promote metastasis by profiling the metastatic cell state of Cdkn2a-deficient cells and dissecting the specific impact of deleting the constituent genes Ink4a and Arf (Aim 1). I will investigate the ability of defined, Cdkn2a-related complex genotypes to dramatically promote metastasis, and I will map metastatic phenotypes to specific gene functions (Aim 2). I will produce tumors with these complex genotypes through the use of Cas12a. I have generated a Cas12a transgenic mouse to take advantage of the unique features of the enzyme, which include the simplicity of targeting multiple loci using a single CRISPR array. Finally, I will explore the metastatic role of the Mtap/Cdkn2a/Cdkn2b locus, which is frequently deleted in its entirety, and test how targeted therapy of Mtap-deficient tumors affects metastatic spread (Aim 3). This work will uncover functional genetic mechanisms by which lung cancer can metastasize, establish critical models for relevant in vivo investigation of metastasis, and identify new therapeutic strategies for treating metastatic lung cancer. Project Number: 1K99CA312806-01 | Fiscal Year: 2026 | NIH Institute/Center: National Cancer Institute (NCI) | Principal Investigator: Jess Hebert | Institution: STANFORD UNIVERSITY, STANFORD, CA | Award Amount: $158,499 | Activity Code: K99 | Study Section: Special Emphasis Panel[ZRG1 CDPT-N (55)] View on NIH RePORTER: https://reporter.nih.gov/project-details/11354122