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Supplementary Methods Figures 1-7 from Metabolic Regulator IAPP (Amylin) Is Required for BRAF and RAS Oncogene-Induced Senescence

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posted on 2023-04-03, 19:49 authored by Sam Garnett, Angeline de Bruyns, Veronique Provencher-Tom, Kendall Dutchak, Ran Shu, David Dankort

Figure S1. Creation of Inducible-BRAF-Inducible Senescent Cells. Figure S2. Summary of lentiviral based shRNA screen for OIS targets. Figure S3. Knockdown of IAPP expression by shRNA. Figure S4. Perturbation of IAPP and ERK2. Figure S5. Loss of IAPP can bypass RAS induced senescence. Figure S6. RAMP3 inhibition affects proliferation arrest. Figure S7. IAPP knockdown cells have a unique profile of glucose metabolites.

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Canadian Institutes of Health Research

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ARTICLE ABSTRACT

Cellular senescence is characterized by a prolonged and predominantly irreversible cell-cycle arrest state, which is linked to loss of tissue function and aging in mammals. Moreover, in response to aberrant oncogenic signals such as those from oncogenic RAS or BRAF, senescence functions as an intrinsic tumor suppressor mechanism restraining tumor progression. In addition to this durable proliferative block, senescent cells adopt altered morphologies, transcriptional profiles, and metabolism, while often possessing unusual heterochromatin formation termed senescence-associated heterochromatic foci. To uncover genes that are required to permit proliferation in the face of sustained oncogene signaling, we conducted an shRNA-based genetic screen in primary cells expressing inducible BRAF. Here we show that depletion of a known glycolysis regulator, islet amylin polypeptide (IAPP also known as amylin), prevents RAS and BRAF oncogene-induced senescence (OIS) in human cells. Importantly, depletion of IAPP resulted in changes of the cells' metabolome and this metabolic reprogramming was associated with widespread alterations in chromatin modifications compared with senescent cells. Conversely, exogenous treatment of IAPP-depleted cells with amylin restored OIS. Together, our results demonstrate that the metabolic regulator IAPP is important regulator of OIS. Moreover, they suggest that IAPP analog treatment or activation of IAPP signaling in RAS/BRAF mutant tumors may have therapeutic potential through senescence induction. These findings demonstrate that IAPP is a novel metabolic regulator of oncogene-induced senescence and use of IAPP analogs may be therapeutically effective to restore growth arrest to BRAF and/or RAS mutant cancers.

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