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In this study, we established a patient\u2010derived xenograft (PDX) model which acquired resistance to the mTOR inhibitor temsirolimus, and explored the underlying mechanisms of resistance acquisition. Temsirolimus was administered to PDX model mice, and one cohort of PDX models acquired resistance after repeated passages. PDX tumors were genetically analyzed by whole\u2010exome sequencing and detected several genetic alterations specific to resistant tumors. Among them, mutations in <jats:italic>ANKRD12<\/jats:italic> and <jats:italic>DNMT1<\/jats:italic> were already identified in the early passage of a resistant PDX model, and we focused on a <jats:italic>DNMT1<\/jats:italic> mutation as a potential candidate for developing the resistant phenotype. While DNMT1 expression in temsirolimus\u2010resistant tumors was comparable with the control tumors, DNMT enzyme activity was decreased in resistant tumors compared with controls. Clustered regularly interspaced short palindromic repeats (CRISPR)\/CRISPR\u2010associated protein 9\u2010mediated heterozygous knockdown of DNMT1 in the temsirolimus\u2010sensitive ccRCC (786\u2010O) cell line was shown to result in a temsirolimus\u2010resistant phenotype in vitro and in vivo. Integrated gene profiles using methylation and microarray analyses of PDX tumors suggested a global shift for the hypomethylation status including promotor regions, and showed the upregulation of several molecules that regulate the mTOR pathway in temsirolimus\u2010resistant tumors. Present study showed the feasibility of PDX model to explore the mechanisms of mTOR resistance acquisition and suggested that genetic alterations, including that of <jats:italic>DNMT1<\/jats:italic>, which alter the methylation status in cancer cells, are one of the potential mechanisms of developing resistance to temsirolimus.<\/jats:p>","DOI":"10.1002\/cam4.3578","type":"journal-article","created":{"date-parts":[[2020,10,27]],"date-time":"2020-10-27T08:19:44Z","timestamp":1603786784000},"page":"119-134","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Functional and genomic characterization of patient\u2010derived xenograft model to study the adaptation to mTORC1 inhibitor in clear cell renal cell 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