1. Genome-wide CRISPR screens in spheroid culture reveal that the tumor suppressor LKB1 inhibits growth via the PIKFYVE lipid kinase.
- Author
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Ferrarone JR, Thomas J, Unni AM, Zheng Y, Nagiec MJ, Gardner EE, Mashadova O, Li K, Koundouros N, Montalbano A, Mustafa M, Cantley LC, Blenis J, Sanjana NE, and Varmus H
- Subjects
- Humans, Cell Proliferation, Cell Line, Tumor, CRISPR-Cas Systems, Lung Neoplasms genetics, Lung Neoplasms pathology, Lung Neoplasms metabolism, Clustered Regularly Interspaced Short Palindromic Repeats genetics, Protein Serine-Threonine Kinases metabolism, Protein Serine-Threonine Kinases genetics, AMP-Activated Protein Kinase Kinases metabolism, AMP-Activated Protein Kinase Kinases genetics, Spheroids, Cellular metabolism, Phosphatidylinositol 3-Kinases metabolism, Phosphatidylinositol 3-Kinases genetics
- Abstract
The tumor suppressor LKB1 is a serine/threonine protein kinase that is frequently mutated in human lung adenocarcinoma (LUAD). LKB1 regulates a complex signaling network that is known to control cell polarity and metabolism; however, the pathways that mediate the tumor-suppressive activity of LKB1 are incompletely defined. To identify mechanisms of LKB1-mediated growth suppression, we developed a spheroid-based cell culture assay to study LKB1-dependent growth. We then performed genome-wide CRISPR screens in spheroidal culture and found that LKB1 suppresses growth, in part, by activating the PIKFYVE lipid kinase. Finally, we used chemical inhibitors and a pH-sensitive reporter to determine that LKB1 impairs growth by promoting the internalization of wild-type EGFR in a PIKFYVE-dependent manner., Competing Interests: Competing interests statement:H.V. is a member of the scientific advisory boards (SABs) for Dragonfly Therapeutics and Surrozen. N.E.S. is an advisor of Vertex and Qiagen and is a co-founder and advisor of OverT Bio. L.C.C. is a co-founder of Faeth Therapeutics, Volastra Therapeutics, and Larkspur Biosciences and is a member of their SABs. L.C.C. is also a co-founder of Agios Pharmaceuticals and is a former member of its SAB and board of directors. M.M. holds stock in 23andMe, Inc. All other authors declare no potential conflicts of interest.
- Published
- 2024
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