Leptin antagonism inhibits prostate cancer xenograft growth and progression

in Endocrine-Related Cancer
View More View Less
  • 1 Australian Prostate Cancer Research Centre - Queensland, Institute of Health and Biomedical Innovation, School of Biomedical Sciences, Queensland University of Technology, Princess Alexandra Hospital, Translational Research Institute, Brisbane, Queensland, Australia
  • 2 Garvan Institute of Medical Research, Sydney, New South Wales, Australia
  • 3 Florey Institute of Neuroscience and Mental Health, University of Melbourne, Melbourne, Victoria, Australia
  • 4 School of Chemistry, University of Melbourne, Melbourne, Victoria, Australia
  • 5 OLPE, LLC, Audubon, Pennsylvania, USA
  • 6 Institute of Medical Microbiology, Semmelweis University, Budapest, Hungary

Correspondence should be addressed to L K Philp: lisa.philp@qut.edu.au
Restricted access

Hyperleptinaemia is a well-established therapeutic side effect of drugs inhibiting the androgen axis in prostate cancer (PCa), including main stay androgen deprivation therapy (ADT) and androgen targeted therapies (ATT). Given significant crossover between the adipokine hormone signalling of leptin and multiple cancer-promoting hallmark pathways, including growth, proliferation, migration, angiogenesis, metabolism and inflammation, targeting the leptin axis is therapeutically appealing, especially in advanced PCa where current therapies fail to be curative. In this study, we uncover leptin as a novel universal target in PCa and are the first to highlight increased intratumoural leptin and leptin receptor (LEPR) expression in PCa cells and patients' tumours exposed to androgen deprivation, as is observed in patients' tumours of metastatic and castrate resistant (CRPC) PCa. We also reveal the world-first preclinical evidence that demonstrates marked efficacy of targeted leptin-signalling blockade, using Allo-aca, a potent, specific, and safe LEPR peptide antagonist. Allo-aca-suppressed tumour growth and delayed progression to CRPC in mice bearing LNCaP xenografts, with reduced tumour vascularity and altered pathways of apoptosis, transcription/translation, and energetics in tumours determined as potential mechanisms underpinning anti-tumour efficacy. We highlight LEPR blockade in combination with androgen axis inhibition represents a promising new therapeutic strategy vital in advanced PCa treatment.

Supplementary Materials

    • Suppl Fig1. Human transcripts (tumour-specific) from Allo-aca (Allo) and Vehicle (Veh) treated tumors cluster separately in PCA plots of normalized CPM of human transcripts, COA plots of normalized CPM of human transcripts and PCA plots of normalized logCPM of differentially expressed (DE) human transcripts. 1448 DE human transcripts were observed with Allo-aca treatment.
    • Suppl Fig2(A-E). Allo-aca efficacy in LNCaP xenograft progression model in castrate Nude mice. Progression over time of (A) tumor xenograft volume from day 0 and (B) serum PSA from castration. Regression over time of (C) tumor volume and (D) serum PSA in the Allo regression cohort n=5). (E) Correlation of endpoint tumor PSA and volume. Cx = castration.
    • Suppl Fig3. Impact of leptin and Allo-aca on LNCaP cell LEPR protein. Representative images of LEPR (green) protein by immunofluorescence in LNCaP cells treated with acute leptin (10 nM) and Allo-aca (100 nM) in vitro under androgen deplete (CSS) or rich (FBS) conditions. Graphically represented as overall LEPR intensity of cells as calculated by Cell Profiler analysis. Statistics: **P≤0.01, ***P≤0.001 vs vehicle; #P≤0.05, ###P≤0.001 vs leptin alone in that condition.
    • Suppl Figure 4 (A-B). Effect of Allo-aca on tumor xenograft phenotype and migration. (A) MetaboAnalyst gene-centric pathway analysis showing significant enrichment of pathways associated with cell adhesion/motility in Allo-aca treated tumours. Analysis was performed on differentially expressed transcripts mapping to the human genome. Pathways are indicated by numbers alongside green circled nodes on the graph and details seen in the corresponding table. Node size corresponds to impact score while node colour corresponds to -log10P significance. (B) Effect of leptin and Allo-aca on androgen-deprived PCa cell transwell migration in vitro. Relative number of migrating LNCaP and C4-2B cells following androgen-deprivation and treatment with leptin doses (0, 2.5, 10 nM), and leptin doses + Allo-aca (1, 10, 100nM); expressed relative to ADT vehicle control. n=3; Statistics: *P<0.05, ***P<0.001, ****P<0.0001, vs ADT vehicle; tP=0.055-0.10, #P<0.05, ##P<0.01, ###P<0.001, vs leptin dose alone.
    • SupplTable1. Top 25 expressed transcripts (based on fpkm value from RNAseq) mapping to the mouse genome expressed in tumour xenografts in Allo-aca (Allo) and Vehicle (Veh) treated mice. Mitochondrial transcripts &#x2013; cell shaded grey. Transcript ID in superscript following gene symbol.

 

Society for Endocrinology

Sept 2018 onwards Past Year Past 30 Days
Abstract Views 219 219 219
Full Text Views 21 21 21
PDF Downloads 33 33 33
  • Bankhead P, Loughrey MB, Fernandez JA, Dombrowski Y, Mcart DG, Dunne PD, Mcquaid S, Gray RT, Murray LJ & Coleman HG et al. 2017 QuPath: open source software for digital pathology image analysis. Scientific Reports 7 16878. (https://doi.org/10.1038/s41598-017-17204-5)

    • Search Google Scholar
    • Export Citation
  • Basaria S, Muller DC, Carducci MA, Egan J & Dobs AS 2006 Hyperglycemia and insulin resistance in men with prostate carcinoma who receive androgen-deprivation therapy. Cancer 106 581588. (https://doi.org/10.1002/cncr.21642)

    • Search Google Scholar
    • Export Citation
  • Cao R, Brakenhielm E, Wahlestedt C, Thyberg J & Cao Y 2001 Leptin induces vascular permeability and synergistically stimulates angiogenesis with FGF-2 and VEGF. PNAS 98 63906395. (https://doi.org/10.1073/pnas.101564798)

    • Search Google Scholar
    • Export Citation
  • Coroniti R, Fario R, Nuno DJ, Otvos L, Scolaro L & Surmacz E 2016 Designer leptin receptor antagonist Allo-aca inhibits VEGF effects in ophthalmic neoangiogenesis models. Frontiers in Molecular Biosciences 3 67. (https://doi.org/10.3389/fmolb.2016.00067)

    • Search Google Scholar
    • Export Citation
  • De Marinis L, , Mancini A, Gentilella R, Perrelli M, Giampietro A, Porcelli T, Tilaro L, Fusco A & Valle D et al. 2004 Growth hormone secretion and leptin in morbid obesity before and after biliopancreatic diversion: relationships with insulin and body composition. Journal of Clinical Endocrinology and Metabolism 89 174180. (https://doi.org/10.1210/jc.2002-021308)

    • Search Google Scholar
    • Export Citation
  • Ettinger SL, Sobel R, Whitmore TG, Akbari M, Bradley DR, Gleave ME & Nelson CC 2004 Dysregulation of sterol response element-binding proteins and downstream effectors in prostate cancer during progression to androgen independence. Cancer Research 64 22122221. (https://doi.org/10.1158/0008-5472.can-2148-2)

    • Search Google Scholar
    • Export Citation
  • Faris JE & Smith MR 2010 Metabolic sequelae associated with androgen deprivation therapy for prostate cancer. Current Opinion in Endocrinology, Diabetes, and Obesity 17 240246. (https://doi.org/10.1097/MED.0b013e3283391fd1)

    • Search Google Scholar
    • Export Citation
  • Ferla R, Bonomi M, Otvos L & Surmacz E 2011 Glioblastoma-derived leptin induces tube formation and growth of endothelial cells: comparison with VEGF effects. BMC Cancer 11 303. (https://doi.org/10.1186/1471-2407-11-303)

    • Search Google Scholar
    • Export Citation
  • Frankenberry KA, Somasundar P, Mcfadden DW & Vona-Davis LC 2004 Leptin induces cell migration and the expression of growth factors in human prostate cancer cells. American Journal of Surgery 188 560565. (https://doi.org/10.1016/j.amjsurg.2004.07.031)

    • Search Google Scholar
    • Export Citation
  • Gonzalez RR, Cherfils S, Escobar M, Yoo JH, Carino C, Styer AK, Sullivan BT, Sakamoto H, Olawaiye A & Serikawa T et al. 2006 Leptin signaling promotes the growth of mammary tumors and increases the expression of vascular endothelial growth factor (VEGF) and its receptor type two (VEGF-R2). Journal of Biological Chemistry 281 2632026328. (https://doi.org/10.1074/jbc.M601991200)

    • Search Google Scholar
    • Export Citation
  • Gorrab A, Pagano A, Ayed K, Chebil M, Derouiche A, Kovacic H & Gati A 2020 Leptin promotes prostate cancer proliferation and migration by stimulating STAT3 pathway Nutrition and Cancer 72 111. (https://doi.org/10.1080/01635581.2020.1792946)

    • Search Google Scholar
    • Export Citation
  • Habib CN, AL-Abd AM, Tolba MF, Khalifa AE, Khedr A, Mosli HA & Abdel-Naim AB 2015 Leptin influences estrogen metabolism and accelerates prostate cell proliferation. Life Sciences 121 1015. (https://doi.org/10.1016/j.lfs.2014.11.007)

    • Search Google Scholar
    • Export Citation
  • Han J & Kaufman RJ 2016 The role of ER stress in lipid metabolism and lipotoxicity. Journal of Lipid Research 57 13291338. (https://doi.org/10.1194/jlr.R067595)

    • Search Google Scholar
    • Export Citation
  • Harris WP, Mostaghel EA, Nelson PS & Montgomery B 2009 Androgen deprivation therapy: progress in understanding mechanisms of resistance and optimizing androgen depletion. Nature Clinical Practice: Urology 6 7685. (https://doi.org/10.1038/ncpuro1296)

    • Search Google Scholar
    • Export Citation
  • Hauck AK & Bernlohr DA 2016 Oxidative stress and lipotoxicity. Journal of Lipid Research 57 19761986. (https://doi.org/10.1194/jlr.R066597)

    • Search Google Scholar
    • Export Citation
  • Horton JD, Goldstein JL & Brown MS 2002 SREBPs: activators of the complete program of cholesterol and fatty acid synthesis in the liver. Journal of Clinical Investigation 109 11251131. (https://doi.org/10.1172/JCI15593)

    • Search Google Scholar
    • Export Citation
  • Jenks MZ, Fairfield HE, Johnson EC, Morrison RF & Muday GK 2017 Sex steroid hormones regulate leptin transcript accumulation and protein secretion in 3T3-L1 cells. Scientific Reports 7 82328232. (https://doi.org/10.1038/s41598-017-07473-5)

    • Search Google Scholar
    • Export Citation
  • Lee S-E, Jang I-S, Park J-S, Lee J-H, Lee S-Y, Baek S-Y, Lee S-H & Lee H 2010 Systemic immunity of obese-diabetes model (db/db) mice. Molecular and Cellular Toxicology 6 143149. (https://doi.org/10.1007/s13273-010-0021-6)

    • Search Google Scholar
    • Export Citation
  • Li Y, Xu S, Mihaylova MM, Zheng B, Hou X, Jiang B, Park O, Luo Z, Lefai E & Shyy JY et al. 2011 AMPK phosphorylates and inhibits SREBP activity to attenuate hepatic steatosis and atherosclerosis in diet-induced insulin-resistant mice. Cell Metabolism 13 376388. (https://doi.org/10.1016/j.cmet.2011.03.009)

    • Search Google Scholar
    • Export Citation
  • Locke JA, Guns ES, Lubik AA, Adomat HH, Hendy SC, Wood CA, Ettinger SL, Gleave ME & Nelson CC 2008 Androgen levels increase by intratumoral de novo steroidogenesis during progression of castration-resistant prostate cancer. Cancer Research 68 64076415. (https://doi.org/10.1158/0008-5472.CAN-07-5997)

    • Search Google Scholar
    • Export Citation
  • Maugham ML, Thomas PB, Crisp GJ, Philp lK, Shah ET, Herington AC, Chen C, Gregory LS, Nelson CC & Seim I et al. 2017 Insights from engraftable immunodeficient mouse models of hyperinsulinaemia. Scientific Reports 7 491 110. (https://doi.org/10.1038/s41598-017-00443-x)

    • Search Google Scholar
    • Export Citation
  • Noda T, Kikugawa T, Tanji N, Miura N, Asai S, Higashiyama S & Yokoyama M 2015 Longterm exposure to leptin enhances the growth of prostate cancer cells. International Journal of Oncology 46 15351542. (https://doi.org/10.3892/ijo.2015.2845)

    • Search Google Scholar
    • Export Citation
  • Otvos Jr L, Kovalszky I, Riolfi M, Ferla R, Olah J, Sztodola A, Nama K, Molino A, Piubello Q & Wade JD et al. 2011 Efficacy of a leptin receptor antagonist peptide in a mouse model of triple-negative breast cancer. European Journal of Cancer 47 15781584. (https://doi.org/10.1016/j.ejca.2011.01.018)

    • Search Google Scholar
    • Export Citation
  • Otvos L, Knappe D, Hoffmann R, Kovalszky I, Olah J, Hewitson TD, Stawikowska R, Stawikowski M, Cudic P & Lin F et al. 2014 Development of second generation peptides modulating cellular adiponectin receptor responses. Frontiers in Chemistry 2 93. (https://doi.org/10.3389/fchem.2014.00093)

    • Search Google Scholar
    • Export Citation
  • Philp LK, Day TK, Butler MS, Laven-Law G, Jindal S, Hickey TE, Scher HI, Butler LM & Tilley WD 2016 Small glutamine-rich tetratricopeptide repeat-containing protein alpha (SGTA) ablation limits offspring viability and growth in mice. Scientific Reports 6 28950. (https://doi.org/10.1038/srep28950)

    • Search Google Scholar
    • Export Citation
  • Philp LK, Muhlhausler BS, Janovska A, Wittert GA, Duffield JA & Mcmillen IC 2008 Maternal overnutrition suppresses the phosphorylation of 5'-AMP-activated protein kinase in liver, but not skeletal muscle, in the fetal and neonatal sheep. American Journal of Physiology: Regulatory, Integrative and Comparative Physiology 295 R1982R1990. (https://doi.org/10.1152/ajpregu.90492.2008)

    • Search Google Scholar
    • Export Citation
  • Philp LK, Rockstroh A, Lehman M, Sadowski MC, Bartonicek N, Wade JD, Otvos L & Nelson CC 2020 Adiponectin receptor activation inhibits prostate cancer xenograft growth endocrine-related cancer. Endocrine-Related Cancer 27 711729.(https://doi.org/10.1530/ERC-20-0297)

    • Search Google Scholar
    • Export Citation
  • Procaccini C, LA Rocca C, Carbone F, De Rosa V, Galgani M & Matarese G 2017 Leptin as immune mediator: interaction between neuroendocrine and immune system. Developmental and Comparative Immunology 66 120129. (https://doi.org/10.1016/j.dci.2016.06.006)

    • Search Google Scholar
    • Export Citation
  • Ramos-Montoya A, Lamb AD, Russell R, Carroll T, Jurmeister S, Galeano-Dalmau N, Massie CE, Boren J, Bon H & Theodorou V et al. 2014 HES6 drives a critical AR transcriptional programme to induce castration-resistant prostate cancer through activation of an E2F1-mediated cell cycle network. EMBO Molecular Medicine 6 651661. (https://doi.org/10.1002/emmm.201303581)

    • Search Google Scholar
    • Export Citation
  • Sadowski MC, Pouwer RH, Gunter JH, Lubik AA, Quinn RJ & Nelson CC 2014 The fatty acid synthase inhibitor triclosan: repurposing an anti-microbial agent for targeting prostate cancer. Oncotarget 5 93629381. (https://doi.org/10.18632/oncotarget.2433)

    • Search Google Scholar
    • Export Citation
  • Sarmento-Cabral A, L-López F & Luque RM 2017 Adipokines and their receptors are widely expressed and distinctly regulated by the metabolic environment in the prostate of male mice: direct role under normal and tumoral conditions. Endocrinology 158 35403552. (https://doi.org/10.1210/en.2017-00370)

    • Search Google Scholar
    • Export Citation
  • Sharifi N 2013 Mechanisms of androgen receptor activation in castration-resistant prostate cancer. Endocrinology 154 40104017. (https://doi.org/10.1210/en.2013-1466)

    • Search Google Scholar
    • Export Citation
  • Shoemaker JE, Lopes TJS, Ghosh S, Matsuoka Y, Kawaoka Y & Kitano H 2012 CTen: a web-based platform for identifying enriched cell types from heterogeneous microarray data. BMC Genomics 13 460. (https://doi.org/10.1186/1471-2164-13-460)

    • Search Google Scholar
    • Export Citation
  • Sieh S, Taubenberger AV, Rizzi SC, Sadowski M, Lehman ML, Rockstroh A, An J, Clements JA, Nelson CC & Hutmacher DW 2012 Phenotypic characterization of prostate cancer LNCaP cells cultured within a bioengineered microenvironment. PLoS ONE 7 e40217. (https://doi.org/10.1371/journal.pone.0040217)

    • Search Google Scholar
    • Export Citation
  • Somasundar P, Frankenberry KA, Skinner H, Vedula G, Mcfadden DW, Riggs D, Jackson B, Vangilder R, Hileman SM & Vona-Davis LC 2004 Prostate cancer cell proliferation is influenced by leptin. Journal of Surgical Research 118 7182. (https://doi.org/10.1016/j.jss.2004.01.017)

    • Search Google Scholar
    • Export Citation
  • Somasundar P, Yu AK, Vona-Davis L & Mcfadden DW 2003 Differential effects of leptin on cancer in vitro. Journal of Surgical Research 113 5055. (https://doi.org/10.1016/s0022-4804(03)00166-5)

    • Search Google Scholar
    • Export Citation
  • Taleb S, Herbin O, Ait-Oufella H, Verreth W, Gourdy P, Barateau V, Merval R, Esposito B, Clement K & Holvoet P et al. 2007 Defective leptin/leptin receptor signaling improves regulatory T cell immune response and protects mice from atherosclerosis. Arteriosclerosis, Thrombosis, and Vascular Biology 27 26912698. (https://doi.org/10.1161/ATVBAHA.107.149567)

    • Search Google Scholar
    • Export Citation
  • Tartaglia LA, Dembski M, Weng X, Deng N, Culpepper J, Devos R, Richards GJ, Campfield LA, Clark FT & Deeds J et al. 1995 Identification and expression cloning of a leptin receptor, OB-R. Cell 83 12631271. (https://doi.org/10.1016/0092-8674(95)90151-5)

    • Search Google Scholar
    • Export Citation
  • Tevz G, Mcgrath S, Demeter R, Magrini V, Jeet V, Rockstroh A, Mcpherson S, Lai J, Bartonicek N & An J et al. 2016 Identification of a novel fusion transcript between human relaxin-1 (RLN1) and human relaxin-2 (RLN2) in prostate cancer. Molecular and Cellular Endocrinology 420 159168. (https://doi.org/10.1016/j.mce.2015.10.011)

    • Search Google Scholar
    • Export Citation
  • Tilki D, Schaeffer EM & Evans CP 2016 Understanding mechanisms of resistance in metastatic castration-resistant prostate cancer: the role of the androgen receptor. European Urology Focus 2 499505. (https://doi.org/10.1016/j.euf.2016.11.013)

    • Search Google Scholar
    • Export Citation
  • Tousignant KD, Rockstroh A, Taherian Fard A, Lehman ML, Wang C, Mcpherson SJ, Philp LK, Bartonicek N, Dinger ME & Nelson CC et al. 2019 Lipid uptake is an androgen-enhanced lipid supply pathway associated with prostate cancer disease progression and bone metastasis. Molecular Cancer Research 17 11661179. (https://doi.org/10.1158/1541-7786.MCR-18-1147)

    • Search Google Scholar
    • Export Citation
  • Vansaun MN 2013 Molecular pathways: adiponectin and leptin signaling in cancer. Clinical Cancer Research 19 19261932. (https://doi.org/10.1158/1078-0432.CCR-12-0930)

    • Search Google Scholar
    • Export Citation
  • Viollet B, Horman S, Leclerc J, Lantier L, Foretz M, Billaud M, Giri S & Andreelli F 2010 AMPK inhibition in health and disease. Critical Reviews in Biochemistry and Molecular Biology 45 276295. (https://doi.org/10.3109/10409238.2010.488215)

    • Search Google Scholar
    • Export Citation
  • Wang H, Burke LJ, Patel J, Tse BWC, Bridle KR, Cogger VC, Li X, Liu X, Yang H & Crawford DHG et al. 2020 Imaging-based vascular-related biomarkers for early detection of acetaminophen-induced liver injury. Theranostics 10 67156727. (https://doi.org/10.7150/thno.44900)

    • Search Google Scholar
    • Export Citation
  • Wek RC 2018 Role of eIF2α kinases in translational control and adaptation to cellular stress. Cold Spring Harbor Perspectives in Biology 10 a032870. (https://doi.org/10.1101/cshperspect.a032870)

    • Search Google Scholar
    • Export Citation
  • Xu CJ, Dong LL, Kang XL, Li ZM & Zhang HY 2020 Leptin promotes proliferation and inhibits apoptosis of prostate cancer cells by regulating ERK1/2 signaling pathway. European Review for Medical and Pharmacological Sciences 24 83418348. (https://doi.org/10.26355/eurrev_202008_22630)

    • Search Google Scholar
    • Export Citation
  • Yang WH, Chang AC, Wang SW, Wang SJ, Chang YS, Chang TM, Hsu SK, Fong YC & Tang CH 2016 Leptin promotes VEGF-C production and induces lymphangiogenesis by suppressing miR-27b in human chondrosarcoma cells. Scientific Reports 6 28647. (https://doi.org/10.1038/srep28647)

    • Search Google Scholar
    • Export Citation
  • Zhu Z, Shi M, Hu W, Estrella H, Engebretsen J, Nichols T, Briere D, Hosea N, Los G & Rejto PA et al. 2012 Dose-dependent effects of small-molecule antagonists on the genomic landscape of androgen receptor binding. BMC Genomics 13 355. (https://doi.org/10.1186/1471-2164-13-355)

    • Search Google Scholar
    • Export Citation