Alternative splice variants of DCLK1 mark cancer stem cells, promote self-renewal and drug-resistance, and can be targeted to inhibit tumorigenesis in kidney cancer

Yang Ge, Nathaniel Weygant, Dongfeng Qu, Randal May, William L. Berry, Jiannan Yao, Parthasarathy Chandrakesan, Wei Zheng, Lichao Zhao, Karena L. Zhao, Michael Drake, Kenneth J Vega, Michael S. Bronze, James J. Tomasek, Guangyu An, Courtney W. Houchen

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Renal cell carcinoma (RCC) is a common and devastating disease characterized by a hypoxic microenvironment, epithelial-mesenchymal transition and potent resistance to therapy evidencing the presence of cancer stem cells (CSCs). Various CSC markers have been studied in RCC, but overall there is limited data on their role and most markers studied have been relatively nonspecific. Doublecortin-like kinase 1 (DCLK1) is a validated CSC marker in the gastrointestinal tract and evidence for an equivalent role in other cancers is accumulating. We used bioinformatics, immunohistochemistry, flow cytometry, spheroid self-renewal and chemoresistance assays in combination with overexpression and siRNA-knockdown to study the stem cell-supportive role of DCLK1 alternative splice variants (DCLK1 ASVs) in RCC. To target tumor cells expressing DCLK1 ASVs directly, we developed a novel monoclonal antibody (CBT-15) and delivered it systemically to RCC tumor xenografts. DCLK1 ASVs were overexpressed, enriched together with CSC markers and predictive of overall and recurrence-free survival in RCC patients. In vitro, DCLK1 ASVs were able to directly stimulate essential molecular and functional characteristics of renal CSCs including expression of aldehyde dehydrogenase, self-renewal and resistance to FDA-approved receptor tyrosine kinase and mTOR inhibitors, while targeted downregulation of DCLK1 reversed these characteristics. Finally, targeting DCLK1 ASV-positive cells with the novel CBT-15 monoclonal antibody blocked RCC tumorigenesis in vivo. These findings establish DCLK1 as a CSC marker with implications for therapy, disease progression and survival in RCC and demonstrate the therapeutic value of DCLK1-targeted monoclonal antibodies against renal CSCs.

Original languageEnglish (US)
Pages (from-to)1162-1175
Number of pages14
JournalInternational Journal of Cancer
Volume143
Issue number5
DOIs
StatePublished - Sep 1 2018
Externally publishedYes

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Neoplastic Stem Cells
Kidney Neoplasms
Drug Resistance
Carcinogenesis
Phosphotransferases
Renal Cell Carcinoma
Monoclonal Antibodies
Cell Self Renewal
Neoplasms
Aldehyde Dehydrogenase
Epithelial-Mesenchymal Transition
Survival
Receptor Protein-Tyrosine Kinases
Computational Biology
Heterografts
Small Interfering RNA
Disease Progression
Gastrointestinal Tract
Flow Cytometry
Stem Cells

Keywords

  • DCLK1
  • cancer stem cell
  • monoclonal antibody therapy
  • renal cancer
  • resistance

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Alternative splice variants of DCLK1 mark cancer stem cells, promote self-renewal and drug-resistance, and can be targeted to inhibit tumorigenesis in kidney cancer. / Ge, Yang; Weygant, Nathaniel; Qu, Dongfeng; May, Randal; Berry, William L.; Yao, Jiannan; Chandrakesan, Parthasarathy; Zheng, Wei; Zhao, Lichao; Zhao, Karena L.; Drake, Michael; Vega, Kenneth J; Bronze, Michael S.; Tomasek, James J.; An, Guangyu; Houchen, Courtney W.

In: International Journal of Cancer, Vol. 143, No. 5, 01.09.2018, p. 1162-1175.

Research output: Contribution to journalArticle

Ge, Y, Weygant, N, Qu, D, May, R, Berry, WL, Yao, J, Chandrakesan, P, Zheng, W, Zhao, L, Zhao, KL, Drake, M, Vega, KJ, Bronze, MS, Tomasek, JJ, An, G & Houchen, CW 2018, 'Alternative splice variants of DCLK1 mark cancer stem cells, promote self-renewal and drug-resistance, and can be targeted to inhibit tumorigenesis in kidney cancer', International Journal of Cancer, vol. 143, no. 5, pp. 1162-1175. https://doi.org/10.1002/ijc.31400
Ge, Yang ; Weygant, Nathaniel ; Qu, Dongfeng ; May, Randal ; Berry, William L. ; Yao, Jiannan ; Chandrakesan, Parthasarathy ; Zheng, Wei ; Zhao, Lichao ; Zhao, Karena L. ; Drake, Michael ; Vega, Kenneth J ; Bronze, Michael S. ; Tomasek, James J. ; An, Guangyu ; Houchen, Courtney W. / Alternative splice variants of DCLK1 mark cancer stem cells, promote self-renewal and drug-resistance, and can be targeted to inhibit tumorigenesis in kidney cancer. In: International Journal of Cancer. 2018 ; Vol. 143, No. 5. pp. 1162-1175.
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abstract = "Renal cell carcinoma (RCC) is a common and devastating disease characterized by a hypoxic microenvironment, epithelial-mesenchymal transition and potent resistance to therapy evidencing the presence of cancer stem cells (CSCs). Various CSC markers have been studied in RCC, but overall there is limited data on their role and most markers studied have been relatively nonspecific. Doublecortin-like kinase 1 (DCLK1) is a validated CSC marker in the gastrointestinal tract and evidence for an equivalent role in other cancers is accumulating. We used bioinformatics, immunohistochemistry, flow cytometry, spheroid self-renewal and chemoresistance assays in combination with overexpression and siRNA-knockdown to study the stem cell-supportive role of DCLK1 alternative splice variants (DCLK1 ASVs) in RCC. To target tumor cells expressing DCLK1 ASVs directly, we developed a novel monoclonal antibody (CBT-15) and delivered it systemically to RCC tumor xenografts. DCLK1 ASVs were overexpressed, enriched together with CSC markers and predictive of overall and recurrence-free survival in RCC patients. In vitro, DCLK1 ASVs were able to directly stimulate essential molecular and functional characteristics of renal CSCs including expression of aldehyde dehydrogenase, self-renewal and resistance to FDA-approved receptor tyrosine kinase and mTOR inhibitors, while targeted downregulation of DCLK1 reversed these characteristics. Finally, targeting DCLK1 ASV-positive cells with the novel CBT-15 monoclonal antibody blocked RCC tumorigenesis in vivo. These findings establish DCLK1 as a CSC marker with implications for therapy, disease progression and survival in RCC and demonstrate the therapeutic value of DCLK1-targeted monoclonal antibodies against renal CSCs.",
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T1 - Alternative splice variants of DCLK1 mark cancer stem cells, promote self-renewal and drug-resistance, and can be targeted to inhibit tumorigenesis in kidney cancer

AU - Ge, Yang

AU - Weygant, Nathaniel

AU - Qu, Dongfeng

AU - May, Randal

AU - Berry, William L.

AU - Yao, Jiannan

AU - Chandrakesan, Parthasarathy

AU - Zheng, Wei

AU - Zhao, Lichao

AU - Zhao, Karena L.

AU - Drake, Michael

AU - Vega, Kenneth J

AU - Bronze, Michael S.

AU - Tomasek, James J.

AU - An, Guangyu

AU - Houchen, Courtney W.

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N2 - Renal cell carcinoma (RCC) is a common and devastating disease characterized by a hypoxic microenvironment, epithelial-mesenchymal transition and potent resistance to therapy evidencing the presence of cancer stem cells (CSCs). Various CSC markers have been studied in RCC, but overall there is limited data on their role and most markers studied have been relatively nonspecific. Doublecortin-like kinase 1 (DCLK1) is a validated CSC marker in the gastrointestinal tract and evidence for an equivalent role in other cancers is accumulating. We used bioinformatics, immunohistochemistry, flow cytometry, spheroid self-renewal and chemoresistance assays in combination with overexpression and siRNA-knockdown to study the stem cell-supportive role of DCLK1 alternative splice variants (DCLK1 ASVs) in RCC. To target tumor cells expressing DCLK1 ASVs directly, we developed a novel monoclonal antibody (CBT-15) and delivered it systemically to RCC tumor xenografts. DCLK1 ASVs were overexpressed, enriched together with CSC markers and predictive of overall and recurrence-free survival in RCC patients. In vitro, DCLK1 ASVs were able to directly stimulate essential molecular and functional characteristics of renal CSCs including expression of aldehyde dehydrogenase, self-renewal and resistance to FDA-approved receptor tyrosine kinase and mTOR inhibitors, while targeted downregulation of DCLK1 reversed these characteristics. Finally, targeting DCLK1 ASV-positive cells with the novel CBT-15 monoclonal antibody blocked RCC tumorigenesis in vivo. These findings establish DCLK1 as a CSC marker with implications for therapy, disease progression and survival in RCC and demonstrate the therapeutic value of DCLK1-targeted monoclonal antibodies against renal CSCs.

AB - Renal cell carcinoma (RCC) is a common and devastating disease characterized by a hypoxic microenvironment, epithelial-mesenchymal transition and potent resistance to therapy evidencing the presence of cancer stem cells (CSCs). Various CSC markers have been studied in RCC, but overall there is limited data on their role and most markers studied have been relatively nonspecific. Doublecortin-like kinase 1 (DCLK1) is a validated CSC marker in the gastrointestinal tract and evidence for an equivalent role in other cancers is accumulating. We used bioinformatics, immunohistochemistry, flow cytometry, spheroid self-renewal and chemoresistance assays in combination with overexpression and siRNA-knockdown to study the stem cell-supportive role of DCLK1 alternative splice variants (DCLK1 ASVs) in RCC. To target tumor cells expressing DCLK1 ASVs directly, we developed a novel monoclonal antibody (CBT-15) and delivered it systemically to RCC tumor xenografts. DCLK1 ASVs were overexpressed, enriched together with CSC markers and predictive of overall and recurrence-free survival in RCC patients. In vitro, DCLK1 ASVs were able to directly stimulate essential molecular and functional characteristics of renal CSCs including expression of aldehyde dehydrogenase, self-renewal and resistance to FDA-approved receptor tyrosine kinase and mTOR inhibitors, while targeted downregulation of DCLK1 reversed these characteristics. Finally, targeting DCLK1 ASV-positive cells with the novel CBT-15 monoclonal antibody blocked RCC tumorigenesis in vivo. These findings establish DCLK1 as a CSC marker with implications for therapy, disease progression and survival in RCC and demonstrate the therapeutic value of DCLK1-targeted monoclonal antibodies against renal CSCs.

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