1 / 41

Targeting Wnt S ignaling in C olorectal C ancer

Targeting Wnt S ignaling in C olorectal C ancer. Özge Keskin Selcuk University , Faculty of Medicine Department of Medical Oncology. Introduction to Wnt signalling. Wnt signaling plays an important role in proliferation , differentiation , motility , survival , apoptosis ,

hisano
Download Presentation

Targeting Wnt S ignaling in C olorectal C ancer

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Targeting WntSignalingin ColorectalCancer Özge Keskin SelcukUniversity, Faculty of Medicine Department of MedicalOncology

  2. IntroductiontoWntsignalling • Wntsignaling plays an important role in • proliferation, • differentiation, • motility, • survival, • apoptosis, • embryonic development, • homeostasis. J. Deitrick and W.M. Pruitt Wnt/β Catenin-Mediated Signaling Commonly Altered in Colorectal Cancer Progress in Molecular Biology and Translational Science, Volume 144. 2016

  3. IntroductiontoWntsignalling • Wnt is a highly insoluble protein ligand which binds to Frizzled (FZD), a cell membrane receptor with seven transmembrane domains. • The insolubility of Wnt is partially caused by the palmitoylation at a conserved cysteine residue, which is a critical posttranslational modification for its signaling ability. J. Deitrick and W.M. Pruitt Wnt/β Catenin-Mediated Signaling Commonly Altered in Colorectal Cancer Progress in Molecular Biology and Translational Science, Volume 144. 2016

  4. IntroductiontoWntsignalling • The WNT family of secreted glycoproteins consists of • WNT1 (INT1), WNT2, WNT2B (WNT13), WNT3 (INT4), WNT3A, WNT4, WNT5A, WNT5B, WNT6, WNT7A, WNT7B, WNT8A, WNT8B, WNT9A (WNT14), WNT9B (WNT14B), WNT10A, WNT10B, WNT11 and WNT16. • WNT signals are transduced through the • Frizzled family comprising seven-transmembrane receptors (FZD1, FZD2, FZD3, FZD4, FZD5, FZD6, FZD7, FZD8, FZD9 and FZD10) and • single-transmembraneco-receptors (LRP5, LRP6, ROR1 and ROR2) to initiate the canonical and non-canonical signaling cascades. KATOH et al. Molecular genetics and targeted therapy of WNT-related human diseases (Review). INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE 40: 587-606, 2017

  5. Wnt controls three major signaling pathways • the Wnt/calcium pathway, • the planar cell polarity pathway, • the canonical Wnt pathway. noncanonical-cateninindependent J. Deitrick and W.M. Pruitt Wnt/β Catenin-Mediated Signaling Commonly Altered in Colorectal Cancer Progress in Molecular Biology and Translational Science, Volume 144. 2016 Novellasdemunt et al. Targeting Wntsignaling in colorectal cancer. A Review in the Theme: Cell Signaling: Proteins, Pathways and Mechanisms. Am J Physiol Cell Physiol 309: C511–C521, 2015.

  6. The Wnt/calcium pathway • Wnt ligands bind to Fz receptors or alternative receptors (RYK or ROR) • increase intracellularCa2+ • activatecalmodulin-dependent proteinkinase II (CaMK2), Jun kinase (JNK), protein kinase (PKC)andcalcineurin • increases the expression of genes associated with cell adhesion and motility J. Deitrick and W.M. Pruitt Wnt/β Catenin-Mediated Signaling Commonly Altered in Colorectal Cancer Progress in Molecular Biology and Translational Science, Volume 144. 2016

  7. The planar cell polarity (PCP)pathway • Regulatescell polarity and morphogenicmovements • Activatessmall GTPases, such as Ras and RhoA(Ras homologous gene family member A),RAC (Ras-related C3 botulinum toxin substrate) and Cdc42 (cell division control protein 42)to activate c-Jun N-terminal kinases. J. Deitrick and W.M. Pruitt Wnt/β Catenin-Mediated Signaling Commonly Altered in Colorectal Cancer Progress in Molecular Biology and Translational Science, Volume 144. 2016 BuketALTINOK, Asuman SUNGUROĞLU. WNT SinyalYolağıveKanser Ankara SağlıkHizmetleriDergisi, Cilt 15, Sayı 2, 2016.

  8. The canonical Wnt pathway • Notonly the most prevalent Wntsignaling pathway but also the most associated with the inititation and progression of cancer. J. Deitrick and W.M. Pruitt Wnt/β Catenin-Mediated Signaling Commonly Altered in Colorectal Cancer Progress in Molecular Biology and Translational Science, Volume 144. 2016

  9. The canonical (beta-catenin-dependent)Wnt pathway • Starts with the binding of Wnt to FZD and co-receptor low-density lipoprotein receptor-related protein 5/6 (LRP5/6) • Terminateswith the transcription of Wnt target genes due to nuclear translocation of β-catenin. In the absence of Wntsignaling, β-catenin is constitutively degraded. J. Deitrick and W.M. Pruitt Wnt/β Catenin-Mediated Signaling Commonly Altered in Colorectal Cancer Progress in Molecular Biology and Translational Science, Volume 144. 2016

  10. Wntsignaling plays a critical role in numerous cellular and developmental processes in normal cells • AberrantWntsignaling is highly associated with numerous cancers,drug resistance and recurrence of tumors. • Overactive Wntsignaling can trigger tumorigenesis in skin, breast, bone marrow, and colon tissue. J. Deitrick and W.M. Pruitt Wnt/β Catenin-Mediated Signaling Commonly Altered in Colorectal Cancer Progress in Molecular Biology and Translational Science, Volume 144. 2016

  11. Wnt pathway is upregulated in both MSI (microsatellite instable)and MSS (microsatellite stable) colorectal cancers. • The great majority (> 90%) of colorectal cancers carry mutations in atleast one Wntsignaling pathway gene • adenomatous polyposis coli (APC) gene • β-catenin (CTNNB1) gene M. Masuda et al. / Pharmacology & Therapeutics 156 (2015) 1–9 Krishnamurthy et al. Targeting the Wnt/beta-catenin pathway in cancer: Update on effectors and inhibitors. Cancer Treatment Reviews 62 (2018) 50–60.

  12. APC mutations • equally occur in replication error (RER)- positive and -negative colorectal cancers • the earliest genetic event during adenoma– carcinoma sequence M. Masuda et al. / Pharmacology & Therapeutics 156 (2015) 1–9

  13. β-catenin (CTNNB1) mutations • Mutuallyexclusiveto APC • About half of colorectal cancers with wild-type APC have mutations in CTNNB1 • β-catenin mutations are seen at a much higher frequency in later stage cancers • Mutatedβ-catenin gene product cannot be properly phosphorylated and is resistant to degradation. M. Masuda et al. / Pharmacology & Therapeutics 156 (2015) 1–9 Sawa et al. Targeting the Wnt signalling pathway in colorectal cancer. Expert Opin. Ther. Targets (2015) 20(4)

  14. Genetic alterations in other Wntsignalling molecules • genesencoding • frizzled 10 (FZD10), • T-cell factors-3 and -4 (TCF3/4) (TCF7L1/2) • axisinhibitor 2 (AXIN2) • APC membranerecruitment protein 1 (AMER1, WTX, or FAM123B) mutated in colorectal cancers. M. Masuda et al. / Pharmacology & Therapeutics 156 (2015) 1–9

  15. Mutations in proteins • Notdirectly involved in the Wntsignaling pathway • May facilitate upregulation of Wnt target genes and play a role in tumorigenesis. • Kirsten rat sarcoma viral oncogene homolog (KRAS) regulates the nuclear localization of β-catenin. • Increased activation of KRAS results in increased nuclear β-catenin. Loss of APC function leads to KRAS oncogene activation promotes β-catenin nuclear translocation J. Deitrick and W.M. Pruitt Wnt/β Catenin-Mediated Signaling Commonly Altered in Colorectal Cancer Progress in Molecular Biology and Translational Science, Volume 144. 2016

  16. Normal intestinal epithelial stem cells reside in the bottom of the crypt intermingled with Paneth cells. Paneth cells constitute the stem cell niche through supply ofWnt3a. Constitutive activation of Wnt signaling by APC or CTNNB1mutation transforms an intestinal epithelial stem cell into adenoma. Adenoma develops into carcinoma through accumulation of additional genetic alterations in the SMAD4, KRAS, and TP53 genes. M. Masuda et al. / Pharmacology & Therapeutics 156 (2015) 1–9

  17. The APC protein forms a multiprotein complex with axin/axin2, casein kinase I α/ε (CKIα/ε), and glycogen synthase kinase 3β (GSK3β), which plays a central role in the degradation of β-catenin  β-catenin destruction complex. Cytoplasmic β-catenin is recruited into the complex phosphorylated and ubiquitinatedwith β-TrCP-containing E3 ubiquitin ligase proteosomal degradation of β-catenin M. Masuda et al. / Pharmacology & Therapeutics 156 (2015) 1–9

  18. Dvl acts as a scaffolding protein to sequester glycogensynthase kinase-3β and axin(β-catenindegradation complex) β-catenin accumulate in the cytosol and translocate to the nucleus function as a transcription coactivator M. Masuda et al. / Pharmacology & Therapeutics 156 (2015) 1–9

  19. 1-Mutations of APC are in the central part of the gene, called the mutation cluster region (MCR: amino acid 1263–1587) truncated APC protein incapable of forming the destruction complex 2-Mutation of the CTNNB1 gene  in the N-terminal phosphorylation sites at Ser45, Thr41, Ser37, and Ser33. M. Masuda et al. / Pharmacology & Therapeutics 156 (2015) 1–9

  20. Surplus β-catenin translocates to the nucleus • coactivator for the T-cell factor (TCF)/lymphoid enhancer factor (LEF) family transcription factors • transactivate a large variety of target genes involved in the proliferation, differentiation, and death of intestinal epithelial cells • c-Jun (JUN) • c-myc (MYC) • cyclin D1 (CCND1) • multidrug resistance 1 (MDR1) (ABCB1) • matrilysin (MMP7) • axin2 (AXIN2) • surviving (BIRC5). M. Masuda et al. / Pharmacology & Therapeutics 156 (2015) 1–9

  21. CYT β-catenin also associates with the coactivatorsp300and CREB-binding protein (CBP). p300  cell differentiation CBP  maintenance of potency. NUC J. Deitrick and W.M. Pruitt Wnt/β Catenin-Mediated Signaling Commonly Altered in Colorectal Cancer Progress in Molecular Biology and Translational Science, Volume 144. 2016

  22. KATOH et al. Molecular genetics and targeted therapy of WNT-related human diseases (Review). INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE 40: 587-606, 2017

  23. KATOH et al. Molecular genetics and targeted therapy of WNT-related human diseases (Review). INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE 40: 587-606, 2017

  24. Anti-WNT signaling therapeutics TargetMechanism of actionDrugStudyphase KATOH et al. Molecular genetics and targeted therapy of WNT-related human diseases (Review). INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE 40: 587-606, 2017

  25. Wnt inhibitors in clinical trials for colorectalcancers Krishnamurthy et al. Targeting the Wnt/beta-catenin pathway in cancer: Update on effectors and inhibitors. Cancer Treatment Reviews 62 (2018) 50–60.

  26. Sawa et al. Targeting the Wnt signalling pathway in colorectal cancer. Expert Opin. Ther. Targets (2015) 20(4)

  27. Wnt-signaling pathway inhibitors in clinical trials for cancer • Inhibitors of the Wnt-Receptor complex • Porcupine inhibitors: Porcupine (PORCN) is a membrane-bound O-acyltransferase (MBOAT), supplies the palmitoyl group to Wnt proteins, a crucial step for Wnt ligand secretion. The Porcupine-selective inhibitor LGK974 blocks Wnt signalling and tumor growth in vivo. • OngoingPhase 1/2 trials with LGK974 in metastatic colorectal cancers with the characteristic mutations like Rnf43/Znrf3, but results are not yet reported. Krishnamurthy et al. Targeting the Wnt/beta-catenin pathway in cancer: Update on effectors and inhibitors. Cancer Treatment Reviews 62 (2018) 50–60.

  28. Inhibitors of the Wnt-Receptor complex • ETC-159 is a small molecule PORCN inhibitor with efficacy inpreclinical models of RSPO-translocated colorectal cancer. Phase I trial since July 2015, with tenpatients in the original cohort (9 Colorectal cancers and 1 renal). There are no responses. Krishnamurthy et al. Targeting the Wnt/beta-catenin pathway in cancer: Update on effectors and inhibitors. Cancer Treatment Reviews 62 (2018) 50–60.

  29. anti-RSPO3 mAb • OMP-131R10 is an anti-RSPO3 mAbthat neutralizes RSPO3 to attenuate canonical WNT signaling through ubiquitylation-mediated FZD degradation. • OMP-131R10 inhibits tumor growth in patient-derived xenograft models of colorectal cancers with RSPO3 fusion. • RSPO translocations are noted in 9% of colorectal cancers. KATOH et al. Molecular genetics and targeted therapy of WNT-related human diseases (Review). INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE 40: 587-606, 2017

  30. Wnt5a mimetics • Foxy-5 is a formulated hexapeptide that can mimic the properties of the Wnt5a molecule to impair cancer cell migration in vitro. • A phase 1 study of Foxy-5 in patients with metastatic colon, breast, and prostate cancer shows no dose limiting toxicity and a phase 1b trial is ongoing. Krishnamurthy et al. Targeting the Wnt/beta-catenin pathway in cancer: Update on effectors and inhibitors. Cancer Treatment Reviews 62 (2018) 50–60.

  31. β-Catenin-destruction complex inhibitors • Tankyrase inhibitors • Tankyrase belongs to the Poly (ADP-ribose) polymerases (PARPs) family. • Two isoforms of Tankyrase, Tankyrase 1 (PARP5a) and Tankyrase 2 (PARP5b) • Increasethe degradation of axin. • Inhibitionof tankyrases leads to stabilization of axins • Tankyraseinhibitor, XAV939 and IWR-1 regulate Axin by inhibiting Tankyrase 1 and Tankyrase 2. Krishnamurthy et al. Targeting the Wnt/beta-catenin pathway in cancer: Update on effectors and inhibitors. Cancer Treatment Reviews 62 (2018) 50–60.

  32. Tankyraseinhibitors • Tankyrase inhibitor, regulate Axin by inhibiting Tankyrase 1 and Tankyrase 2. • Mouse tumor xenografts and patient-derived sphere cultures of colorectal cancer were incubated with a Tankyrase inhibitor, NVP-TNKS656 in addition to AKT and PI3K inhibitors. A high nuclear beta-catenin level predicted for apoptosis with NVPTNKS656 in combination with PI3K and AKT inhibitors suggesting the tankyrase inhibitor could overcome resistance to these inhibitors. • XAV939 inhibits colony formation by APC-deficient colorectal cancer cells and is thus considered a promising molecular therapeutic drug. • Another tankyrase 1/2 inhibitor, JW55 (Tocris Bioscience, UK), has been reported to reduce the growth of SW480 colorectal cancer cells harboring APC mutation. Sawa et al. Targeting the Wnt signalling pathway in colorectal cancer. Expert Opin. Ther. Targets (2015) 20(4)

  33. Interaction of β-cateninand CBP • Stabilized nuclear β-catenin associates with BCL9, CBP, p300, EZH2 and SMARCA4 to activate transcription of TCF/LEF-target genes. β-catenin inhibitors have been developed with a focus on its protein-protein interactions. • PRI-724,targetedthe interaction ofβ-catenin andCBP. KATOH et al. Molecular genetics and targeted therapy of WNT-related human diseases (Review). INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE 40: 587-606, 2017. Bahrami et al. Therapeutic Potential of Targeting Wnt/b-Catenin Pathway in Treatment of Colorectal Cancer: Rational and Progress Journal of Cellular Biochemistry 118:1979–1983 (2017)

  34. PRI-724 in advancedsolid tumors • Adverse events: Grade 2 diarrhea, bilirubin elevation, hypophosphatemia, nausea, fatigue, anorexia, thrombocytopenia,alkalinephosphatase elevation.Grade 3: hyperbilirubinemia • Phase2 trial in metastatic colorectalcancer. • PRI-724+FOLFOX6+bevacizumab • Not yetrecruitment (NCT02413853) M. Masuda et al. / Pharmacology & Therapeutics 156 (2015) 1–9 J ClinOncol 31, 2013 (suppl; abstr 2501)

  35. Monoclonal antibody against frizzled receptors • Wnt proteins exert their biological activities by binding to the transmembrane cell-surface G-protein-coupled FZD receptors. • There are 10 FZD receptors in the human genome. • OMP-18R5(vanituctumab) is a human monoclonal antibody that was originallyintended to target the FZD7 receptor but was later found to bind tofive FZD receptors (1, 2, 5, 7, and 8) . • OMP-18R5 binds to the FZD receptors and inhibits their ligand binding and Wntsignaling. M. Masuda et al. / Pharmacology & Therapeutics 156 (2015) 1–9

  36. Monoclonal antibody against frizzled receptors • In the majority of colorectal cancers, β-catenin destruction complex is not properly formed due to truncation of APC, and only the molecules downstream of APC can be considered as targets for Wnt signal blockage. • FZD receptors transduceWntsignalling upstream of APC. • OMP-18R5inhibited the growth of xenografted human colorectal cancer with wild-type APC and CTNNB1, but not that of colorectal cancer harboring APC or CTNNB1 gene mutations. M. Masuda et al. / Pharmacology & Therapeutics 156 (2015) 1–9

  37. TNIK as a regulator of Wnt signalling • TNIK was originally identified as the novel Germinal CenterKinase(GCK) family, a subgroup of the STE20 kinase family • Interactwith tumor necrosis factor (TNF)-receptor- associated factor 2 (TRAF2) and NCK adaptor protein (NCK1). • TNIK disrupted the actin cytoskeleton and thereby inhibited cell spreading. M. Masuda et al. / Pharmacology & Therapeutics 156 (2015) 1–9

  38. TNIK as a regulator of Wnt signalling • Because the great majority of colorectal cancers carry mutations ineither APC or CTNNB1, only Wnt signal molecules downstream of the β-catenin destruction complex should be considered as therapeutic targets. • TNIK is an essential regulator of the β-catenin and TCF4 transcription complex, the most downstream component of Wntsignaling. M. Masuda et al. / Pharmacology & Therapeutics 156 (2015) 1–9

  39. The crystal structure of TNIK with PD407824, aWee1Chk1 Inhibitor also inhibits TNIK. • ON 108600, is a dual inhibitor of CK2 and TNIK. • An aminothiazolebased TNIK inhibitor NCB-0001 was identified as a potent TNIK inhibitor. M. Masuda et al. / Pharmacology & Therapeutics 156 (2015) 1–9

  40. Thanksforyourattentıon…..

More Related