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The essential clathrin adaptor protein complex-2 is tumor suppressive specifically in vivo [CRISPR Screen]

ABSTRACT: The essential clathrin adaptor protein complex-2 is tumor suppressive specifically in vivo [CRISPR Screen]

PROVIDER: PRJNA1224449 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR32362904.fastq.gz	Fastqsanger.gz
	SRR32362905.fastq.gz	Fastqsanger.gz
	SRR32362906.fastq.gz	Fastqsanger.gz
	SRR32362907.fastq.gz	Fastqsanger.gz
	SRR32362908.fastq.gz	Fastqsanger.gz

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The essential clathrin adaptor protein complex-2 is tumor suppressive specifically in vivo

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Project description:The microenvironment is a rich source of new cancer targets. While whole-genome CRISPR/Cas9 screens have identified vulnerabilities in cultured cells, it is a challenge to use this approach in tumors to interrogate the microenvironment. To this end, we screened a panel of pancreatic cancer lines grown in culture versus as tumors with a phosphatidylinositol metabolism CRISPR/Cas9 library. This revealed the Adaptor Protein complex-2 (AP2) of clathrin-mediated endocytosis behaves as common essential in culture but tumor suppressive in vivo. These polar-opposite phenotypes were attributed to different microenvironments. In culture, AP2 loss reduces endocytosis of transferrin and iron import. In tumors, alternative iron transport pathways allow pro-tumor signaling to manifest. In the most sensitive case, AP2 loss reprograms the plasma membrane proteome, retaining integrins on the surface leading to downregulation of the tumor-suppressor PTEN. Analysis of human tumors suggests a similar tumor suppressive activity. AP2 may foreshadow other proteins unique to the tumor-microenvironment.

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The essential clathrin adapter protein complex-2 is tumor suppressive specifically in vivo.

Project description:The microenvironment is a rich source of new cancer targets. We thus used a targeted single-guide RNA library to screen a panel of human pancreatic cancer lines for genes uniquely affecting tumorigenesis. Here we show inactivation of the Adapter Protein complex-2 of clathrin-mediated endocytosis reduces cell growth in vitro, but completely oppositely, promotes tumor growth in vivo. In culture, loss of the complex reduces transferrin endocytosis and iron import required for cell fitness. In tumors, alternative iron transport pathways allow pro-tumor effects of Adapter Protein complex-2 loss to manifest. In the most sensitive case, this is attributed to reprogramming the plasma membrane proteome, retaining integrins on the surface leading to Focal Adhesion Kinase phosphorylation and induction of proliferative signals. Adapter Protein complex-2 function in tumorigenesis is thus dependent upon the microenvironment, behaving as a common essential gene in culture via iron import, but as a tumor suppressor in tumors via integrin trafficking.

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