Figure 1.
Strategy to identify mechanistic determinants of NEPC by SB mutagenesis. Shown is the overall strategy as described in the text. Step 1: SB mutagenesis was done on the NPp53 mice GEEM of prostate cancer. NPp53-SB(−) and NPp53-SB(+) mice were generated, and phenotypic and molecular analyses were performed. Step 2: Transcriptomic data from SB mice were integrated with comparable data from human prostate cancer patient to assess conservation. Step 3: CIS of transposon integration were determined, and CIS genes were identified. Step 4: CIS genes were prioritized as candidate mechanistic determinants of NEPC by integrating transcriptomic and genomic data from mouse and human prostate cancer. Step 5: Candidates were functionally validated in human prostate cancer and mouse NEPC organoid models. Refer to the image caption for details. The illustration shows a multi-step research strategy to identify mechanistic determinants of neuroendocrine prostate cancer (NEPC) using Sleeping Beauty (SB) mutagenesis. Step 1 shows the initial step involving SB mutagenesis on NPp53 mice, generating NPp53-SB(minus) and NPp53-SB(plus) mice, followed by phenotypic and molecular analyses of mouse SB tumors. Step 2 depicts transcriptomic analyses, including differential gene expression analysis, master regulator analysis, and OncoMatch of mouse SB tumors to NEPC patients, highlighting cross-species analysis of RNA seq data. Step 3 focuses on genomic analyses, identifying common insertion site (CIS) genes enriched in NEPC through the identification of CIS genes, mapping CIS to associated genes, and differential expression of CIS genes in NEPC. Step 4 illustrates the prioritization of CIS genes as candidate drivers of NEPC through cross-species data integration, involving CIS genes differentially expressed in NEPC, master regulators of NEPC, and CINDy to identify modulators of NEPC. Step 5 shows the functional validation of candidate CIS genes, indicating that gain of function of SIRT1 promotes NEPC, loss of function of SIRT1 suppresses NEPC, and pharmacological inhibition of SIRT1 inhibits NEPC.

Strategy to identify mechanistic determinants of NEPC by SB mutagenesis. Shown is the overall strategy as described in the text. Step 1: SB mutagenesis was done on the NPp53 mice GEEM of prostate cancer. NPp53-SB(−) and NPp53-SB(+) mice were generated, and phenotypic and molecular analyses were performed. Step 2: Transcriptomic data from SB mice were integrated with comparable data from human prostate cancer patient to assess conservation. Step 3: CIS of transposon integration were determined, and CIS genes were identified. Step 4: CIS genes were prioritized as candidate mechanistic determinants of NEPC by integrating transcriptomic and genomic data from mouse and human prostate cancer. Step 5: Candidates were functionally validated in human prostate cancer and mouse NEPC organoid models.

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