Single-Cell-Derived Primary Rectal Carcinoma Cell Lines Reflect Intratumor Heterogeneity Associated with Treatment Response

Rüdiger Braun, Lena Anthuber, Daniela Hirsch, Darawalee Wangsa, Justin Lack, Nicole E. McNeil, Kerstin Heselmeyer-Haddad, Irianna Torres, Danny Wangsa, Markus A. Brown, Anthony Tubbs, Noam Auslander, E. Michael Gertz, Philip R. Brauer, Margaret C. Cam, Dan L. Sackett, Jens K. Habermann, Andre Nussenzweig, Eytan Ruppin, Zhongqiu ZhangDaniel W. Rosenberg, Thomas Ried


PURPOSE: The standard treatment of patients with locally advanced rectal cancer consists of preoperative chemoradiotherapy (CRT) followed by surgery. However, the response of individual tumors to CRT is extremely diverse, presenting a clinical dilemma. This broad variability in treatment response is likely attributable to intratumor heterogeneity (ITH). EXPERIMENTAL DESIGN: We addressed the impact of ITH on response to CRT by establishing single-cell-derived cell lines (SCDCL) from a treatment-naïve rectal cancer biopsy after xenografting. RESULTS: Individual SCDCLs derived from the same tumor responded profoundly different to CRT in vitro. Clonal reconstruction of the tumor and derived cell lines based on whole-exome sequencing revealed nine separate clusters with distinct proportions in the SCDCLs. Missense mutations in SV2A and ZWINT were clonal in the resistant SCDCL, but not detected in the sensitive SCDCL. Single-cell genetic analysis by multiplex FISH revealed the expansion of a clone with a loss of PIK3CA in the resistant SCDCL. Gene expression profiling by tRNA-sequencing identified the activation of the Wnt, Akt, and Hedgehog signaling pathways in the resistant SCDCLs. Wnt pathway activation in the resistant SCDCLs was confirmed using a reporter assay. CONCLUSIONS: Our model system of patient-derived SCDCLs provides evidence for the critical role of ITH for treatment response in patients with rectal cancer and shows that distinct genetic aberration profiles are associated with treatment response. We identified specific pathways as the molecular basis of treatment response of individual clones, which could be targeted in resistant subclones of a heterogenous tumor.

Original languageEnglish
JournalClinical cancer research : an official journal of the American Association for Cancer Research
Issue number13
Pages (from-to)3468-3480
Number of pages13
Publication statusPublished - 01.07.2020

Research Areas and Centers

  • Research Area: Luebeck Integrated Oncology Network (LION)


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