DOCSLIB.ORG

Supplementary Data

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Supplementary Data SUPPLEMENTARY DATA Supplementary Figure 1. Cytokine-Cytokine receptor interaction pathway components predicted to be regulated by let-7 miRNAs. Pathway analysis performed using DIANA LAB mirPATH v.3.(1) Predicted let-7 targets are indicated by red star symbol. ©2017 American Diabetes Association. Published online at http://diabetes.diabetesjournals.org/lookup/suppl/doi:10.2337/db16-1405/-/DC1 SUPPLEMENTARY DATA Supplementary Figure 2. TNF-alpha signaling pathway components predicted to be regulated by let-7 miRNAs. Pathway analysis performed using DIANA LAB mirPATH v.3.(1) Predicted let-7 targets are indicated by red star symbol. ©2017 American Diabetes Association. Published online at http://diabetes.diabetesjournals.org/lookup/suppl/doi:10.2337/db16-1405/-/DC1 SUPPLEMENTARY DATA Supplementary Figure 3. NF-Kappa-B signaling pathway components predicted to be regulated by let-7 miRNAs. Pathway analysis performed using DIANA LAB mirPATH v.3.(1) Predicted let-7 targets are indicated by red star symbol ©2017 American Diabetes Association. Published online at http://diabetes.diabetesjournals.org/lookup/suppl/doi:10.2337/db16-1405/-/DC1 SUPPLEMENTARY DATA Supplementary Figure 4. Bioinformatic analysis of human and mouse Lin28b 3’UTR sequences for interaction with miRNA. Multiple let-7 recognition sites are present. Targetscan (www.targetscan.org). Human LIN28B ENST00000345080.4 3' UTR length: 4549 Mouse LIN28B ENST00000345080.4 3' UTR length: 4430 ©2017 American Diabetes Association. Published online at http://diabetes.diabetesjournals.org/lookup/suppl/doi:10.2337/db16-1405/-/DC1 SUPPLEMENTARY DATA Supplementary Figure 5. Lin28b gene expression in carotid plaque tissue from non-diabetic and diabetic patients (n=4-5, ± SEM). Expression was normalized to 18S for gene expression analysis. *P ≤ 0.05. LIN28B 40 NS 30 20 10 Relative expression qPCR expression Relative 0 c tic ti e e b b ia a Di n-D o N ©2017 American Diabetes Association. Published online at http://diabetes.diabetesjournals.org/lookup/suppl/doi:10.2337/db16-1405/-/DC1 SUPPLEMENTARY DATA Supplementary Figure 6. Lipoxins can restore let-7 levels in SMCs and ECs. Let-7d expression in SMCs pretreated with the ALX/FPR2 antagonist BOC2 (10uM, 30 minutes) followed by LXA 4 (1nM, 30 minutes) and TNF-α (1 ng/ml; 24 hours) stimulation. Expression was normalized to U87 for miRNA analysis (n=3, ± SEM). *P ≤ 0.05. Let-7d 7 * 6 5 4 NS 2 1 Relative expression qPCR expression Relative 0 d O C A e O LX F LX N B T treat DMS A + F Un TN A + LX + BOC F N T ©2017 American Diabetes Association. Published online at http://diabetes.diabetesjournals.org/lookup/suppl/doi:10.2337/db16-1405/-/DC1 SUPPLEMENTARY DATA Supplementary Figure 7. Expression levels of genes associated with ECM and fibrosis in aortic tissue from C57BL/6 mice that received a single tail-vein injection of let-7d mimic or miR-NC for 24 hours (n=6, ± SEM). Expression was normalized to 18S for gene expression analysis and U87 for miRNA analysis. *P ≤ 0.05. 4 miR-NC Let-7d mimic 3 2 * 1 Relative expression qPCR expression Relative 0 1 AI1 A1 MP2 P 3 4A M MMP9 MMP12 COL1A1 COL COL ©2017 American Diabetes Association. Published online at http://diabetes.diabetesjournals.org/lookup/suppl/doi:10.2337/db16-1405/-/DC1 SUPPLEMENTARY DATA Supplementary Figure 8. Flow diagram outlining the methodology for assessing secreted cytokine responses from human carotid plaque biopsies following transfection with scrambled miRNA mimic or let-7d mimic for 24 hours. Human Plaque Studies Carotid Endarterectomy (Recruited from St. Vincent’s University Hospital, Dublin) Plaque - Histological assessment Plaque dissection, Culturing, Transfection (24 hours) Let-7d mimic delivery (Untreated, Control mimic, Let-7d mimic) RNA extraction – Collect plaque assess let-7d levels in supernatant – each plaque secreted proteome (confirm transfection) analysis Let-7d Pooling Strategy: 50 * Scr 40 30 • Pool 1 (Scrambled mimic): 300ul per 20 plaque (1200ul final volume). 10 Let-7d Relative expression qPCR expression Relative 0 • Pool 2 (Let-7d mimic): 300ul per plaque (1200ul final volume. 7d mimic Let- Scrambled mimic ©2017 American Diabetes Association. Published online at http://diabetes.diabetesjournals.org/lookup/suppl/doi:10.2337/db16-1405/-/DC1 SUPPLEMENTARY DATA Supplementary Figure 9. Transfection of human carotid plaque explants with let-7d mimic modulates inflammatory cytokine release. (A) Raw dot-blots indicating cytokine levels in supernatant collected from human carotid plaques 24 hours after transfection with let-7d mimic or scrambled miRNA mimic (n=4). (B) Information on cytokines measured in Proteome Profiler Human XL Cytokine Array. A. B. Coordinate Analyte Entrez Coordinate Analyte Entrez Coordinate Analyte Entrez A1, A2 Reference D9, D10 IFN-γ G9, G10 MCP-3 A3, A4 Adiponectin D11, D12 IGFBP-2 G11, G12 M-CSF A5, A6 ApoA1 D13, D14 IGFBP-3 G13, G14 MIF Scrambled A7, A8 Angiogenin D15, D16 IL-1α G15, G16 MIG A9, A10 ANGPT1 D17, D18 IL-1β G17, G18 MIP-1α/MIP-1β Control A11, A12 ANGPT2 D19, D20 IL-1ra G19, G20 MIP-3α A13, A14 TNFSF13B D21, D22 IL-2 G21, G22 MIP-3β miRNA A15, A16 BDNF D23, D24 IL-3 G23, G24 MMP-9 A17, A18 C5/C5a E1, E2 IL-4 H1, H2 Myeloperoxidase A19, A20 CD14 E3, E4 IL-5 H3, H4 Osteopontin A21, A22 TNFRSF8 E5, E6 IL-6 H5, H6 PDGF-AA A23, A24 Reference E7, E8 IL-8 H7, H8 PDGF-AB/BB B3, B4 TNFSF5, E9, E10 IL-10 H9, H10 Pentraxin B5, B6 CHI3L1, E11, E12 IL-11 H11, H12 PF4 B7, B8 CFD E13, E14 IL-12 H13, H14 RAGE B9, B10 C-Reactive E15, E16 IL-13 H15, H16 RANTES B11, B12 CRP E17, E18 IL-15 H17, H18 RBP-4 B13, B14 Cystatin E19, E20 IL-16 H19, H20 Relaxin-2 Let-7d B15, B16 Dkk-1 E21, E22 IL-17A H21, H22 Resistin miRNA B17, B18 DPPIV E23, E24 IL-18 H23, H24 SDF-1α B19, B20 EGF F1, F2 IL-19 I1, I2 Serpin B21, B22 Emmprin F3, F4 IL-22 I3, I4 SHBG C3, C4 ENA-78 F5, F6 IL-23 I5, I6 ST2 C5, C6 Endoglin F7, F8 IL-24 I7, I8 TARC C7, C8 CD178, F9, F10 IL-27 I9, I10 TFF3 C9, C10 FGF-2 F11, F12 IL-31 I11, I12 TfR C11, C12 FGF-7 F13, F14 IL-32 I13, I14 TGF-α C13, C14 FGF-19 F15, F16 IL-33 I15, I16 Thrombospondin-1 C15, C16 Flt-3 F17, F18 IL-34 I17, I18 TNF-α C17, C18 G-CSF F19, F20 IP-10 I19, I20 uPAR C19, C20 GDF-15 F21, F22 I-TAC I21, I22 VEGF C21, C22 GM-CSF F23, F24 KLK3 J1, J2 Reference D1, D2 GROα G1, G2 Leptin J5, J6 VDBP D3, D4 Somatotropin G3, G4 LIF J7, J8 CD31 D5, D6 HGF G5, G6 Lipocalin-2 J9, J10 TIM-3 D7, D8 ICAM-1 G7, G8 MCP-1 J11, J12 VCAM-1 J23, J24 Negative Control ©2017 American Diabetes Association. Published online at http://diabetes.diabetesjournals.org/lookup/suppl/doi:10.2337/db16-1405/-/DC1 SUPPLEMENTARY DATA Supplementary Table 1. In silico prediction of let-7-transcript interactions. Predicted Let-7 target? Position in the context++ context++ score Pct** GENE (Yes/No) miRNA UTR seed match score* percentile** * IL1B NO MCP1 NO P21 NO P53 NO PCNA NO PDGFR NO ICAM1 NO VCAM1 NO LIN28B mmu-let-7a-1-3p 2337-2343 7mer-1A -0.17 95 N/A LIN28B mmu-let-7a-2-3p 1945-1951 7mer-1A -0.05 61 N/A LIN28B mmu-let-7a-5p 36-43 8mer -0.4 93 0.95 LIN28B mmu-let-7a-5p 2409-2415 7mer-m8 -0.35 91 0.91 LIN28B mmu-let-7a-5p 2581-2587 7mer-m8 -0.21 73 0.91 LIN28B mmu-let-7a-5p 3997-4003 7mer-m8 -0.31 87 0.86 LIN28B mmu-let-7a-5p 4185-4191 7mer-1A -0.15 61 0.37 YES LIN28B mmu-let-7b-3p 2337-2343 7mer-1A -0.15 93 N/A LIN28B mmu-let-7b-5p 36-43 8mer -0.4 93 0.95 LIN28B mmu-let-7b-5p 2409-2415 7mer-m8 -0.32 88 0.91 LIN28B mmu-let-7b-5p 2581-2587 7mer-m8 -0.23 75 0.91 LIN28B mmu-let-7b-5p 3997-4003 7mer-m8 -0.31 87 0.86 LIN28B mmu-let-7b-5p 4185-4191 7mer-1A -0.17 64 0.37 LIN28B mmu-let-7c-1-3p 1944-1951 8mer -0.03 49 N/A ©2017 American Diabetes Association. Published online at http://diabetes.diabetesjournals.org/lookup/suppl/doi:10.2337/db16-1405/-/DC1 SUPPLEMENTARY DATA Position in the context++ context++ score Pct** GENE miRNA UTR seed match score* percentile** * LIN28B mmu-let-7c-2-3p 2337-2343 7mer-1A -0.17 95 N/A LIN28B mmu-let-7c-5p 36-43 8mer -0.4 93 0.95 LIN28B mmu-let-7c-5p 2409-2415 7mer-m8 -0.34 90 0.91 LIN28B mmu-let-7c-5p 2581-2587 7mer-m8 -0.21 73 0.91 LIN28B mmu-let-7c-5p 3997-4003 7mer-m8 -0.31 87 0.86 LIN28B mmu-let-7c-5p 4185-4191 7mer-1A -0.15 61 0.37 LIN28B mmu-let-7d-5p 36-43 8mer -0.41 94 0.95 LIN28B mmu-let-7d-5p 2409-2415 7mer-m8 -0.32 87 0.91 LIN28B mmu-let-7d-5p 2581-2587 7mer-m8 -0.2 68 0.91 LIN28B mmu-let-7d-5p 3997-4003 7mer-m8 -0.32 87 0.86 LIN28B mmu-let-7d-5p 4185-4191 7mer-1A -0.19 65 0.37 LIN28B mmu-let-7e-5p 36-43 8mer -0.4 93 0.95 LIN28B mmu-let-7e-5p 2409-2415 7mer-m8 -0.35 91 0.91 LIN28B mmu-let-7e-5p 2581-2587 7mer-m8 -0.21 73 0.91 LIN28B mmu-let-7e-5p 3997-4003 7mer-m8 -0.31 86 0.86 LIN28B mmu-let-7e-5p 4185-4191 7mer-1A -0.15 61 0.37 LIN28B mmu-let-7f-1-3p 2337-2343 7mer-1A -0.12 89 N/A LIN28B mmu-let-7f-2-3p 2337-2343 7mer-1A -0.09 83 N/A LIN28B mmu-let-7f-5p 36-43 8mer -0.4 93 0.95 LIN28B mmu-let-7f-5p 2409-2415 7mer-m8 -0.35 91 0.91 LIN28B mmu-let-7f-5p 2581-2587 7mer-m8 -0.21 73 0.91 LIN28B mmu-let-7f-5p 3997-4003 7mer-m8 -0.31 86 0.86 LIN28B mmu-let-7f-5p 4185-4191 7mer-1A -0.15 61 0.37 LIN28B mmu-let-7g-3p 2649-2655 7mer-1A -0.06 42 N/A LIN28B mmu-let-7g-5p 36-43 8mer -0.4 93 0.95 LIN28B mmu-let-7g-5p 2409-2415 7mer-m8 -0.33 89 0.91 ©2017 American Diabetes Association.
Load more

Recommended publications
  • An Ontogenetic Switch Drives the Positive and Negative Selection of B Cells
    An ontogenetic switch drives the positive and negative selection of B cells Xijin Xua, Mukta Deobagkar-Lelea, Katherine R. Bulla, Tanya L. Crockforda, Adam J. Meadb, Adam P. Cribbsc, David Simsc, Consuelo Anzilottia, and Richard J. Cornalla,1 aMedical Research Council Human Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, OX3 9DS Oxford, United Kingdom; bMedical Research Council Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, OX3 9DS Oxford, United Kingdom; and cMedical Research Council, Weatherall Institute of Molecular Medicine, Centre for Computational Biology, Weatherall Institute of Molecular Medicine, University of Oxford, OX3 9DS Oxford, United Kingdom Edited by Michael Reth, University of Freiburg, Freiburg, Germany, and approved January 6, 2020 (received for review September 3, 2019) + Developing B cells can be positively or negatively selected by self- BM HSCs increased CD5 B-1a B cell development (15), while antigens, but the mechanisms that determine these outcomes are expression of let-7b in FL pro-B cells blocked the development of incompletely understood. Here, we show that a B cell intrinsic B-1 B cells (17). These findings support the notion of hard-wired switch between positive and negative selection during ontogeny differences during ontogeny, but possibly downstream of the HSC is determined by a change from Lin28b to let-7 gene expression. commitment stage. Ectopic expression of a Lin28b transgene in murine B cells restored Several lines of evidence also suggest that B-1 B cells can un- the positive selection of autoreactive B-1 B cells by self-antigen in dergo positive selection, which is linked to their B cell receptor adult bone marrow.
    [Show full text]
  • The LIN28B-IMP1 Post-Transcriptional Regulon Has Opposing Effects On
    Washington University School of Medicine Digital Commons@Becker Open Access Publications 2018 The IN28BL -IMP1 post-transcriptional regulon has opposing effects on oncogenic signaling in the intestine Blair B. Madison et al Follow this and additional works at: https://digitalcommons.wustl.edu/open_access_pubs Downloaded from genesdev.cshlp.org on August 7, 2018 - Published by Cold Spring Harbor Laboratory Press The LIN28B–IMP1 post-transcriptional regulon has opposing effects on oncogenic signaling in the intestine Priya Chatterji,1,2,14 Kathryn E. Hamilton,1,3,14 Shun Liang,4 Sarah F. Andres,1 H.R. Sagara Wijeratne,4 Rei Mizuno,1 Lauren A. Simon,1,3 Philip D. Hicks,1 Shawn W. Foley,5 Jason R. Pitarresi,1 Andres J. Klein-Szanto,6,7 Amanda T. Mah,8 Laurianne Van Landeghem,9 Brian D. Gregory,5 Christopher J. Lengner,10 Blair B. Madison,11 Premal Shah,4,12 and Anil K. Rustgi1,2,13 1Department of Medicine, Division of Gastroenterology, 2Department of Genetics, University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania 19014, USA; 3Department of Pediatrics, Division of Gastroenterology, Children’s Hospital of Philadelphia, University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania 19014, USA; 4Department of Genetics, Rutgers University, New Brunswick, New Jersey 08901, USA; 5Department of Biology, University of Pennsylvania, Philadelphia, Pennsylvania 19014, USA; 6Department of Pathology, 7Cancer Biology Program, Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111, USA; 8Department of Medicine, Hematology Division, Stanford University, Stanford, California 94305, USA; 9Department of Molecular Biomedical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina 27607, USA; 10Department of Biomedical Sciences, School of Veterinary Medicine, Institute for Regenerative Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA; 11Department of Medicine, Division of Gastroenterology, Washington University School of Medicine, St.
    [Show full text]
  • LIN28B Promotes the Development of Neuroendocrine Prostate Cancer
    LIN28B promotes the development of neuroendocrine prostate cancer Jessica M. Lovnicki, … , Martin Gleave, Xuesen Dong J Clin Invest. 2020. https://doi.org/10.1172/JCI135373. Research In-Press Preview Oncology Graphical abstract Find the latest version: https://jci.me/135373/pdf LIN28B promotes the development of neuroendocrine prostate cancer Jessica Lovnicki1, *, Yu Gan1, 2, *, Tingting Feng1, 3, Yinan Li1, Ning Xie1, Chia-Hao Ho1, Ahn Lee1, Xufeng Chen4, Lucia Nappi1, Bo Han3, Ladan Fazli1, Jiaoti Huang4, Martin Gleave1, Xuesen Dong1 1) The Vancouver Prostate Centre, Department of Urologic Sciences, University of British Columbia, Vancouver, British Columbia, Canada 2) Department of Urology, Xiangya Hospital, Central South University, Changsha, Hunan Province, China 3) The Key Laboratory of Experimental Teratology, Ministry of Education, and Department of Pathology, School of Basic Medical Sciences, Shandong University, Jinan, Shandong Province, China 4) Department of Pathology, Duke University School of Medicine, Durham, North Carolina, USA *) The authors equally contribute to this work. Corresponding author: Xuesen Dong, Ph.D. 2660 Oak Street, Vancouver, British Columbia, Canada V6H 3Z6, Tel: 604-875-4111 Fax: 604-875-5654 Email: [email protected] Disclosure Statement: The authors have declared that no conflict of interest exists. 1 Abstract Therapy-induced neuroendocrine prostate cancer (t-NEPC) is a highly aggressive subtype of prostate cancer with poor patient survival. Emerging evidence indicates that t-NEPC can develop when prostate adenocarcinoma cells acquire cancer stem-like cell signaling in the presence of androgen receptor inhibition, followed by re-differentiation toward neuroendocrine lineage and subsequent t-NEPC progression. Whether the stem-like signaling is controlled by the core pluripotency stem cell genes (e.g., LIN28 and SOX2) remains unknown.
    [Show full text]
  • Detailed Characterization of Human Induced Pluripotent Stem Cells Manufactured for Therapeutic Applications
    Stem Cell Rev and Rep DOI 10.1007/s12015-016-9662-8 Detailed Characterization of Human Induced Pluripotent Stem Cells Manufactured for Therapeutic Applications Behnam Ahmadian Baghbaderani 1 & Adhikarla Syama2 & Renuka Sivapatham3 & Ying Pei4 & Odity Mukherjee2 & Thomas Fellner1 & Xianmin Zeng3,4 & Mahendra S. Rao5,6 # The Author(s) 2016. This article is published with open access at Springerlink.com Abstract We have recently described manufacturing of hu- help determine which set of tests will be most useful in mon- man induced pluripotent stem cells (iPSC) master cell banks itoring the cells and establishing criteria for discarding a line. (MCB) generated by a clinically compliant process using cord blood as a starting material (Baghbaderani et al. in Stem Cell Keywords Induced pluripotent stem cells . Embryonic stem Reports, 5(4), 647–659, 2015). In this manuscript, we de- cells . Manufacturing . cGMP . Consent . Markers scribe the detailed characterization of the two iPSC clones generated using this process, including whole genome se- quencing (WGS), microarray, and comparative genomic hy- Introduction bridization (aCGH) single nucleotide polymorphism (SNP) analysis. We compare their profiles with a proposed calibra- Induced pluripotent stem cells (iPSCs) are akin to embryonic tion material and with a reporter subclone and lines made by a stem cells (ESC) [2] in their developmental potential, but dif- similar process from different donors. We believe that iPSCs fer from ESC in the starting cell used and the requirement of a are likely to be used to make multiple clinical products. We set of proteins to induce pluripotency [3]. Although function- further believe that the lines used as input material will be used ally identical, iPSCs may differ from ESC in subtle ways, at different sites and, given their immortal status, will be used including in their epigenetic profile, exposure to the environ- for many years or even decades.
    [Show full text]
  • RNA-Binding Protein LIN28B Inhibits Apoptosis Through Regulation of the AKT2/FOXO3A/BIM Axis in Ovarian Cancer Cells
    Signal Transduction and Targeted Therapy www.nature.com/sigtrans ARTICLE OPEN RNA-binding protein LIN28B inhibits apoptosis through regulation of the AKT2/FOXO3A/BIM axis in ovarian cancer cells Xiaojuan Lin1,2, Jianfeng Shen 1, Dan Peng3, Xinhong He1,4, Congjian Xu5, Xiaojun Chen5, Janos L. Tanyi6, Kathleen Montone7,YiFan8, Qihong Huang9,10,11, Lin Zhang1,6 and Xiaomin Zhong 3 LIN28B is an evolutionarily conserved RNA-binding protein that regulates mRNA translation and miRNA let-7 maturation in embryonic stem cells and developing tissues. Increasing evidence demonstrates that LIN28B is activated in cancer and serves as a critical oncogene. However, the underlying molecular mechanisms of LIN28B function in tumorigenesis are still largely unknown. Here we report that LIN28B was expressed in over half of the patients with epithelial ovarian cancer who were examined (n = 584). Functional experiments demonstrated that LIN28B inhibited ovarian cancer cell apoptosis. Furthermore, we showed that the proapoptotic factor BIM played an essential role in the antiapoptotic function of LIN28B. RNA-IP microarray analysis suggested that LIN28B binds to mRNAs that are associated with the DNA damage pathway, such as AKT2, in ovarian cancer cells. By binding to AKT2 mRNA and enhancing its protein expression, LIN28B regulated FOXO3A protein phosphorylation and decreased the transcriptional level of BIM, which antagonized the antiapoptosis activity of LIN28B. Taken together, these results mechanistically linked LIN28B and the AKT2/FOXO3A/BIM axis to the apoptosis
    [Show full text]
  • The LIN28B/Let-7 Axis Is a Novel Therapeutic Pathway in Multiple Myeloma
    The LIN28B/let-7 axis is a novel therapeutic pathway in Multiple Myeloma The Harvard community has made this article openly available. Please share how this access benefits you. Your story matters Citation Manier, S., J. T. Powers, A. Sacco, S. V. Glavey, D. Huynh, M. R. Reagan, K. Z. Salem, et al. 2016. “The LIN28B/let-7 axis is a novel therapeutic pathway in Multiple Myeloma.” Leukemia 31 (4): 853-860. doi:10.1038/leu.2016.296. http://dx.doi.org/10.1038/ leu.2016.296. Published Version doi:10.1038/leu.2016.296 Citable link http://nrs.harvard.edu/urn-3:HUL.InstRepos:32630451 Terms of Use This article was downloaded from Harvard University’s DASH repository, and is made available under the terms and conditions applicable to Other Posted Material, as set forth at http:// nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of- use#LAA HHS Public Access Author manuscript Author ManuscriptAuthor Manuscript Author Leukemia Manuscript Author . Author manuscript; Manuscript Author available in PMC 2017 April 24. Published in final edited form as: Leukemia. 2017 April ; 31(4): 853–860. doi:10.1038/leu.2016.296. The LIN28B/let-7 axis is a novel therapeutic pathway in Multiple Myeloma Salomon Manier1,2,3, John T. Powers4, Antonio Sacco1, Siobhan V. Glavey1, Daisy Huynh1, Michaela R. Reagan1, Karma Z. Salem1, Michele Moschetta1, Jiantao Shi1, Yuji Mishima1, Catherine Roche-Lestienne3, Xavier Leleu2, Aldo M. Roccaro1, George Q. Daley4, and Irene M. Ghobrial1 1Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston 02215 MA, USA 2Service des Maladies du Sang, CHRU Lille, 59000 Lille, France 3Jean-Pierre Aubert Research Centre, INSERM U1172, University Lille 2, 59000 Lille, France 4Division of Pediatric Hematology/Oncology, Children’s Hospital, Harvard Medical School, Boston 02215 MA, USA Abstract MYC is a major oncogenic driver of Multiple Myeloma (MM) and yet almost no therapeutic agents exist that target MYC in MM.
    [Show full text]
  • Lin28b, LET-7, and the Molecular Pathogenesis of Colon Cancer
    University of Pennsylvania ScholarlyCommons Publicly Accessible Penn Dissertations Fall 2010 Lin28B, LET-7, and the Molecular Pathogenesis of Colon Cancer Catrina E. King University of Pennsylvania, [email protected] Follow this and additional works at: https://repository.upenn.edu/edissertations Part of the Cancer Biology Commons Recommended Citation King, Catrina E., "Lin28B, LET-7, and the Molecular Pathogenesis of Colon Cancer" (2010). Publicly Accessible Penn Dissertations. 1551. https://repository.upenn.edu/edissertations/1551 This paper is posted at ScholarlyCommons. https://repository.upenn.edu/edissertations/1551 For more information, please contact [email protected]. Lin28B, LET-7, and the Molecular Pathogenesis of Colon Cancer Abstract Lin28b is an RNA-binding protein that inhibits biogenesis of tumor-suppressive microRNAs of the let-7 family, and is involved in induction of pluripotency. Although LIN28B has been implicated in cancer, a specific oler in colon tumorigenesis has not been elucidated. We have determined that colon tumors exhibit decreased levels of mature let-7 isoforms, and that constitutive let-7 expression inhibits migration and invasion of colon cancer cells in vitro. Importantly, down-regulation of let-7a and let-7b in colon tumors correlates with increased expression of LIN28B, suggesting tumor-promoting properties of this let-7 inhibitor. In order to determine the role of LIN28B in colon cancer, we constitutively expressed LIN28B in immortalized human colonic epithelial cells and colon cancer cell lines via retroviral transduction. We found that constitutive LIN28B expression promotes migration, invasion, and soft-agar colony formation in vitro, as well as differentiated and metastatic phenotypes in vivo. Additionally, constitutive LIN28B expression modulates levels of several mRNA transcripts including the established let-7 targets IGF2BP1 and HMGA2.
    [Show full text]
  • LIN28B Fosters Colon Cancer Migration, Invasion and Transformation Through Let-7-Dependent and -Independent Mechanisms
    Oncogene (2011) 30, 4185–4193 & 2011 Macmillan Publishers Limited All rights reserved 0950-9232/11 www.nature.com/onc ORIGINAL ARTICLE LIN28B fosters colon cancer migration, invasion and transformation through let-7-dependent and -independent mechanisms CE King1,3,LWang1,RWinograd1,BBMadison1,3,PSMongroo1,3, CN Johnstone1 and AK Rustgi1,2,3 1Gastroenterology Division and Department of Medicine, University of Pennsylvania, Philadelphia, PA, USA; 2Department of Genetics, University of Pennsylvania, Philadelphia, PA, USA and 3Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA, USA Lin28b is an RNA-binding protein that inhibits biogenesis as inhibition of let-7 microRNA biogenesis is a cardinal of let-7 microRNAs. LIN28B is overexpressed in diverse feature of their functions. cancers, yet a specific role in the molecular pathogenesis The let-7 family of microRNAs comprises isoforms of colon cancer has to be elucidated. We have determined with highly conserved sequences that exhibit functional that human colon tumors exhibit decreased levels of redundancy (Pasquinelli et al., 2000; Zhao et al., 2010). mature let-7 isoforms and increased expression of Let-7 biogenesis resembles that of most microRNAs, in LIN28B. To determine LIN28B’s mechanistic role in which the various isoforms are transcribed initially by colon cancer, we expressed LIN28B in immortalized RNA polymerase II as pri-microRNAs, and processed colonic epithelial cells and human colon cancer cell lines. by Drosha and DGCR8 into pre-microRNAs that are We found that LIN28B promotes cell migration, invasion subsequently exported from the nucleus (Lee et al., and transforms immortalized colonic epithelial cells. 2002, 2003; Gregory et al., 2004; Han et al., 2004).
    [Show full text]
  • A Variant in LIN28B Is Associated with 2D:4D Finger-Length Ratio, a Putative Retrospective Biomarker of Prenatal Testosterone Exposure
    ARTICLE A Variant in LIN28B Is Associated with 2D:4D Finger-Length Ratio, a Putative Retrospective Biomarker of Prenatal Testosterone Exposure Sarah E. Medland,1,9 Tetyana Zayats,2,3,9 Beate Glaser,2,3 Dale R. Nyholt,1 Scott D. Gordon,1 Margaret J. Wright,1 Grant W. Montgomery,1 Megan J. Campbell,1 Anjali K. Henders,1 Nicholas J. Timpson,2,3 Leena Peltonen,4,5,6,7 Dieter Wolke,8 Susan M. Ring,3 Panos Deloukas,4 Nicholas G. Martin,1 George Davey Smith,2,3 and David M. Evans2,3,* The ratio of the lengths of an individual’s second to fourth digit (2D:4D) is commonly used as a noninvasive retrospective biomarker for prenatal androgen exposure. In order to identify the genetic determinants of 2D:4D, we applied a genome-wide association approach to 1507 11-year-old children from the Avon Longitudinal Study of Parents and Children (ALSPAC) in whom 2D:4D ratio had been measured, as well as a sample of 1382 12- to 16-year-olds from the Brisbane Adolescent Twin Study. A meta-analysis of the two scans identified a single variant in the LIN28B gene that was strongly associated with 2D:4D (rs314277: p ¼ 4.1 3 10À8) and was subsequently independently replicated in an additional 3659 children from the ALSPAC cohort (p ¼ 1.53 3 10À6). The minor allele of the rs314277 variant has previously been linked to increased height and delayed age at menarche, but in our study it was associated with increased 2D:4D in the direction opposite to that of previous reports on the correlation between 2D:4D and age at menarche.
    [Show full text]
  • LIN28B Antibody A
    Revision 1 C 0 2 - t LIN28B Antibody a e r o t S Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) 6 9 Web: [email protected] 1 www.cellsignal.com 4 # 3 Trask Lane Danvers Massachusetts 01923 USA For Research Use Only. Not For Use In Diagnostic Procedures. Applications: Reactivity: Sensitivity: MW (kDa): Source: UniProt ID: Entrez-Gene Id: WB, IP H Endogenous 32, 21 Rabbit Q6ZN17 389421 Product Usage Information Application Dilution Western Blotting 1:1000 Immunoprecipitation 1:50 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody. Specificity / Sensitivity LIN28B Antibody detects endogenous levels of total LIN28B protein. Species Reactivity: Human Source / Purification Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino acid residues at the carboxyl terminus of human LIN28B. Antibodies are purified by Protein A and peptide affinity chromatography. Background LIN28A and LIN28B are conserved, developmentally regulated RNA binding proteins that inhibit the processing and maturation of the let-7 family of miRNAs (1,2). The let-7 miRNAs have been implicated in repression of oncogenes such as Ras, Myc, and HMGA2 (3). It has recently been shown that upregulation of LIN28A and LIN28B in primary human tumors and human cancer cell lines is correlated with downregulation of let-7 miRNAs (4). LIN28 genes are reported to be involved in primordial germ cell development and germ cell malignancy (5). In addition, allelic variation in LIN28B is associated with regulating the timing of puberty in humans (6).
    [Show full text]
  • LIN28B Induces a Differentiation Program Through CDX2 in Colon Cancer
    LIN28B induces a differentiation program through CDX2 in colon cancer Kensuke Suzuki, … , Michael P Verzi, Anil K. Rustgi JCI Insight. 2021. https://doi.org/10.1172/jci.insight.140382. Research In-Press Preview Cell biology Gastroenterology Graphical abstract Find the latest version: https://jci.me/140382/pdf Title LIN28B induces a differentiation program through CDX2 in colon cancer Authors and affiliations Kensuke Suzuki, 1 Yasunori Masuike,1 Rei Mizuno,2 Uma M. Sachdeva, 3 Priya Chatterji,4 Sarah F. Andres,5 Wenping Sun,6 Andres J. Klein-Szanto,7 Sepideh Besharati,8 Helen E. Remotti,8 Michael P. Verzi,9 Anil K. Rustgi1* 1 Herbert Irving Comprehensive Cancer Center, Division of Digestive and Liver Disease, Department of Medicine, Columbia University Irving Medical Center, New York, New York, USA.2 Department of Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan. .3 Division of Thoracic Surgery, Massachusetts General Hospital, Boston, Massachusetts. 4 Broad Institute of Massachusetts Institute of Technology and Harvard University, Cambridge, Massachusetts, USA.5 Papé Family Pediatric Research Institute, Oregon Health & Science University, Portland, Oregon, USA.6 Institute for Biomedical informatics, University of Pennsylvania, Philadelphia, Pennsylvania, USA.7 Histopathology Facility and Cancer Biology Program, Fox Chase Cancer Center, Philadelphia, Pennsylvania, USA. 8 Department of Pathology & Cell Biology, Columbia University Medical Center, New York, New York, USA. 9 Department of genetics, Human Genetics Institute of New Jersey, Rutgers University, Piscataway, New Jersey, USA * Corresponding Author: Anil K. Rustgi, MD Irving Professor of Medicine Director, Herbert Irving Comprehensive Cancer Center Columbia University Medical Center 1 1130 St. Nicholas Avenue, New York, New York, 10032 Phone: 2157716361 Email: [email protected] RUNNING TITLE: LIN28B-CDX2-AMACR axis in colon cancer differentiation Conflict of interest statement The authors have declared that no conflict of interest exists.
    [Show full text]
  • LIN28B Induces a Differentiation Program Through CDX2 in Colon Cancer
    LIN28B induces a differentiation program through CDX2 in colon cancer Kensuke Suzuki, … , Michael P. Verzi, Anil K. Rustgi JCI Insight. 2021;6(9):e140382. https://doi.org/10.1172/jci.insight.140382. Research Article Cell biology Gastroenterology Graphical abstract Find the latest version: https://jci.me/140382/pdf RESEARCH ARTICLE LIN28B induces a differentiation program through CDX2 in colon cancer Kensuke Suzuki,1 Yasunori Masuike,1 Rei Mizuno,2 Uma M. Sachdeva,3 Priya Chatterji,4 Sarah F. Andres,5 Wenping Sun,6 Andres J. Klein-Szanto,7 Sepideh Besharati,8 Helen E. Remotti,8 Michael P. Verzi,9 and Anil K. Rustgi1 1Herbert Irving Comprehensive Cancer Center, Division of Digestive and Liver Disease, Department of Medicine, Vagelos College of Physicians and Surgeons, Columbia University Irving Medical Center, New York, New York, USA. 2Department of Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan. 3Division of Thoracic Surgery, Massachusetts General Hospital, Boston, Massachusetts, USA. 4Broad Institute of MIT and Harvard University, Cambridge, Massachusetts, USA. 5Papé Family Pediatric Research Institute, Oregon Health & Science University, Portland, Oregon, USA. 6Institute for Biomedical informatics, University of Pennsylvania, Philadelphia, Pennsylvania, USA. 7Histopathology Facility and Cancer Biology Program, Fox Chase Cancer Center, Philadelphia, Pennsylvania, USA. 8Department of Pathology and Cell Biology, Columbia University Medical Center, New York, New York, USA. 9Department of Genetics, Human Genetics Institute of New Jersey, Rutgers University, Piscataway, New Jersey, USA. Most colorectal cancers (CRCs) are moderately differentiated or well differentiated, a status that is preserved even in metastatic tumors. However, the molecular mechanisms underlying CRC differentiation remain to be elucidated. Herein, we unravel a potentially novel posttranscriptional regulatory mechanism via a LIN28B/CDX2 signaling axis that plays a critical role in mediating CRC differentiation.
    [Show full text]