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PTP1B Deficiency Enables the Ability of a High-Fat Diet to Drive the Invasive Character of PTEN-Deficient Prostate Cancers

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PTP1B Deficiency Enables the Ability of a High-Fat Diet to Drive the Invasive Character of PTEN-Deficient Prostate Cancers Published OnlineFirst March 28, 2016; DOI: 10.1158/0008-5472.CAN-15-1501 Cancer Priority Report Research PTP1B Deficiency Enables the Ability of a High-Fat Diet to Drive the Invasive Character of PTEN-Deficient Prostate Cancers David P. Labbe1,2, Noriko Uetani1,Valerie Vinette1,3, Laurent Lessard4, Isabelle Aubry1, Eva Migon1, Jacinthe Sirois1, Jody J. Haigh5, Louis R. Begin 6, Lloyd C. Trotman7, Marilene Paquet8, and Michel L. Tremblay1,2,3 Abstract Diet affects the risk and progression of prostate cancer, but vation, interpreted to reflect a heightened sensitivity to IGF-1 the interplay between diet and genetic alterations in this disease stimulation upon HFD feeding. Prostate-specific overexpres- is not understood. Here we present genetic evidence in the sion of PTP1B was not sufficienttoinitiateprostatecancer, mouse showing that prostate cancer progression driven by arguingthatitactedasadiet-dependentmodifier of prostate À À loss of the tumor suppressor Pten is mainly unresponsive to cancer development in Pten / mice. Our findings offer a a high-fat diet (HFD), but that coordinate loss of the protein preclinical rationale to investigate the anticancer effects of tyrosine phosphatase Ptpn1 (encoding PTP1B) enables a highly PTP1B inhibitors currently being studied clinically for diabetes À À À À invasive disease. Prostate cancer in Pten / Ptpn1 / mice treatment as a new modality for management of prostate was characterized by increased cell proliferation and Akt acti- cancer. Cancer Res; 76(11); 3130–5. Ó2016 AACR. Introduction metabolism and cancer and is now a validated therapeutic target for diabetes, obesity, and breast cancer (7). Prostate cancer is the most frequently diagnosed cancer in The promise of PTP1B-directed therapeutics prompted us to North American men and is the second leading cause of can- further characterize the role of PTP1B in prostate cancer initiation cer-related deaths (1). Lifetime prostate cancer risk is overwhelm- and progression using preclinical models. Here we report that ingly associated with environmental factors (2) such as diet (3) prostate-specific overexpression of PTP1B does not lead to pros- and obesity (4). However, the mechanistic links remain elusive. tate transformation, ruling out the possibility that PTP1B alone is We recently described the protein tyrosine phosphatase 1B capable of inducing prostate cancer. Surprisingly, however, com- (PTP1B; encoded by PTPN1) as an androgen receptor (AR)- À À À À pound Ptpn1 / ; PtenPE / (PE, prostate epithelium) mice show a regulated phosphatase, which plays a tumor-promoting role in significant increase in prostate cancer invasiveness, but only when prostate cancer (5), and showed that the PTPN1 gene is coam- these mice are fed a high-fat diet (HFD), seemingly through the plified with the AR in metastatic prostate cancer (6). PTP1B is enhancement of insulin-like growth factor 1 (IGF-1)-mediated located at a critical node in signaling pathways that regulate Akt activation. Together with the observation that the levels of PTP1B protein are consistently increased following Pten loss, these 1Goodman Cancer Research Centre, McGill University, Montreal, results suggest that PTP1B acts as a diet-dependent tumor sup- Quebec, Canada. 2Division of Experimental Medicine, Department of pressor in the context of prostate cancer that is driven by the Medicine, McGill University, Montreal, Quebec, Canada. 3Department absence of Pten. Importantly, our results highlight that cancer of Biochemistry, McGill University, Montreal, Quebec, Canada. 4 progression can be altered by a synergistic cooperation between Research Group in Molecular Oncology and Endocrinology, Depart- fi ment of Medical Biology, UniversiteduQu ebec a Trois-Rivieres, Trois- genetic and environmental factors. Finally, these ndings indicate Rivieres, Quebec, Canada. 5Mammalian Functional Genetics Labora- that diet and levels of PTP1B enzymatic activity are important tory, Division of Blood Cancers, Australian Centre for Blood Diseases, parameters when considering the clinical use of PTP1B-targeted Department of Clinical Haematology, Monash University and Alfred Health Alfred Centre, Melbourne,Victoria, Australia. 6Service d'anato- therapeutics. mopathologie, Hopital^ du Sacre-C œur de Montreal, Montreal, Quebec, Canada. 7Cold Spring Harbor Laboratory, Cold Spring Harbor, New York. 8FacultedeM edecine Vet erinaire, Departement de Pathologie et Materials and Methods de Microbiologie, Universite de Montreal, St-Hyacinthe, Quebec, Animal husbandry Canada. Genotypes for Ptpn1, Pten,andPB-Cre4 were determined Note: Supplementary data for this article are available at Cancer Research by PCR (Supplementary Table S1). C57BL/6 mice were fed Online (http://cancerres.aacrjournals.org/). either regular lab chow (Harlan Laboratories, #2920X), or a Corresponding Author: Michel L. Tremblay, McGill University, Goodman Cancer HFD (Harlan Laboratories, #TD.07011) from the time of wean- Research Centre, 1160 Pine Avenue West, Room 617, Montreal, Quebec H3A 1A3, ing.Ouranimalprotocolfollowed the ethical guidelines Canada. Phone: 514-398-7290; Fax: 514-398-6769; E-mail: of the Canadian Council on Animal Care, and was approved [email protected] by the McGill University Research and Ethics Animal Commit- doi: 10.1158/0008-5472.CAN-15-1501 tee. Detailed are reported in Supplementary Materials and Ó2016 American Association for Cancer Research. Methods. 3130 Cancer Res; 76(11) June 1, 2016 Downloaded from cancerres.aacrjournals.org on September 24, 2021. © 2016 American Association for Cancer Research. Published OnlineFirst March 28, 2016; DOI: 10.1158/0008-5472.CAN-15-1501 PTP1B Is a Diet-Dependent Tumor Suppressor in Prostate Cancer Generation of PTPN1 knock-in mice Analyses of protein expression We used Gateway-compatible Rosa26 locus targeting vectors to Protein expression was analyzed as described previously (5). generate a Cre/loxP conditional Rosa26-targeted transgenic mouse Additional details can be found in Supplementary Materials and that overexpresses human PTP1B (Fig. 1A; ref. 8). The detailed Methods. procedure is described in Supplementary Materials and Methods. Histopathologic and immunohistochemical analyses Tissue processing/histopathologic examination were done Live imaging according to conventional procedures, as reported in Supplemen- fl Mice were anesthetized with 2-chloro-2-(di uoromethoxy)- tary Materials and Methods. 1,1,1-trifluoro-ethane (isoflurane), injected intraperitoneally with 50 mL luciferin (Caliper Life Sciences), and imaged with the Cell culture and lentiviral infections þ þ À À use of the IVIS Spectrum preclinical in vivo imaging system (Perkin Immortalized Ptpn1 / and Ptpn1 / mouse embryonic fibro- Elmer), according to the manufacturer's instructions. blasts (MEF; ref. 9) were infected with a lentiviral short hairpin Figure 1. PTP1B overexpression in the prostatic epithelium does not drive cancer. A, Rosa26-targeted (R26) pCAGG-promoter-based construct drives strong expression of 3X- PTPN1/WT FLAG-PTP1B protein (R26 ), together with an eGFP/luciferase reporter in the prostate PE, following breeding with a PB-Cre4 mouse. B and C, validation of the R26PTPN1/WT transgene by live imaging (B) and FLAG immunoprecipitation (C). D, representative photomicrographs of H&E staining to show DLP cystic dilation (left; stars), normal epithelium in the AP of R26WT/WT mice (middle; arrows) along with some regions of focal hyperplasia in R26PTPN1/WT mice (middle; arrow) and lymphocytic infiltration in VP (right; arrows). www.aacrjournals.org Cancer Res; 76(11) June 1, 2016 3131 Downloaded from cancerres.aacrjournals.org on September 24, 2021. © 2016 American Association for Cancer Research. Published OnlineFirst March 28, 2016; DOI: 10.1158/0008-5472.CAN-15-1501 Labbe et al. RNA vector against Pten (shPten) or a scramble sequence prostate (DLP; Fig. 1D; Supplementary Table S2). Together, these (kindly provided by Nicholas R. Leslie, Heriot Watt University, results suggest that, unlike breast cancer, prostate-specificPTP1B Edinburgh, United Kingdom). Complementary details can be overexpression in intact murine prostates is insufficient to initiate found in Supplementary Materials and Methods. prostate cancer. To confirm that PTP1B is instead required for prostate cancer progression, as we previously documented in cell-based and Statistical analyses À À À À xenograft assays (5), we generated Ptpn1 / ; PtenPE / compound Statistical analyses were carried out with use of GraphPad Prism mice. Homozygous inactivation of Pten in the PE driven by PB- 6.0 Software. Cre4 is a robust prostate cancer model that yields mPIN by the age of 6 weeks. Following prostate-specific Pten inactivation, histo- Results pathologic analysis revealed no differences in the number (per- þ þ To assess whether PTP1B has a prostate cancer–initiating role centage) of prostate ductules affected by mPIN between Ptpn1 / À À similar to what we previously observed in breast cancer (10), we and Ptpn1 / mice (Supplementary Fig. S1A and S1B). Likewise, À À À À þ þ generated a Cre/loxP conditional Rosa26-targeted transgenic mouse genetic ablation of Ptpn1 / per se (Ptpn1 / ; PtenPE / mice) did PTPN1/WT that overexpresses a FLAG-PTPN1 transgene (R26 ), togeth- not lead to prostate gland transformation, nor to any significant er with an eGFP/luciferase reporter (Fig. 1A). Prostate-specific alterations beyond mPIN in the AP and ventral prostates (VP) of PTP1B overexpression was achieved by crossing this model with all 6-week-old mice (Supplementary
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