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Cha Kuin Taiteit Het Wat U U W Tuin CHA KUIN TAITEIT US009868792B2HET WAT UU W TUIN (12 ) United States Patent ( 10 ) Patent No. : US 9 , 868 ,792 B2 Zabel et al. ( 45 ) Date of Patent: Jan . 16 , 2018 (54 ) METHODS OF ENHANCING ANTI- TUMOR OTHER PUBLICATIONS IMMUNITY BY ADMINISTERING ANTIBODIES TO THE CCRL2 CHEMERIN Pachynski et al. , “ The chemoattractant chemerin suppresses mela RECEPTOR noma by recruiting natural killer cell antitumor defenses” , J Exp Med . , Jul. 2 , 2012 , pp . 1427 - 1435 , 209 ( 8 ), The Rockefeller Uni versity Press , New York , NY. (71 ) Applicant: The Board of Trustees of the Leland Wittamer et al. , “ Specific Recruitment of Antigen - presenting Cells Stanford Junior University , Stanford , by Chemerin , a Novel Processed Ligand from Human Inflammatory CA (US ) Fluids ” , J Exp Med ., Oct . 6 , 2003 , pp . 977 - 985 , 198 ( 7 ) , The Rockefeller University Press , New York , NY. (72 ) Inventors : Brian A . Zabel , Redwood City , CA Zabel et al . , “ Chemerin activation by serine proteases of the (US ) ; Eugene C . Butcher , Portola coagulation , fibrinolytic , and inflammatory cascades” , · J Biol Valley , CA (US ) ; Russell K . Chem . , Oct. 14 , 2005 , pp . 34661 - 34666 , 280 ( 41) , American Society Pachynski, Saint Louis, MO (US ) ; for Biochemistry and Molecular Biology , Rockville , MD . Justin Monnier , San Francisco , CA Zabel et al ., Mast cell -expressed orphan receptor CCRL2 binds (US ) chemerin and is required for optimal induction of IgE -mediated passive cutaneous anaphylaxis : , J Exp Med ., Sep . 15 , 2008 , pp . ( 73 ) Assignee : The Board of Trustees of the Leland 2207 -2220 , 205 ( 10 ) , The Rockefeller University Press, New York , NY Stanford Junior University, Stanford , Monnier et al ., “ Expression , regulation , and function of atypical CA (US ) chemerin receptor CCRL2 on endothelial cells ” , J Immunol . , Jun . 13 , 2012 , pp . 956 - 967 , 189 ( 2 ) , The American Association of Immu ( * ) Notice : Subject to any disclaimer , the term of this nologists , Inc ., Rockville , MD . patent is extended or adjusted under 35 Gonzalvo - Feo et al. , “ Endothelial cell -derived chemerin promotes U . S . C . 154 ( b ) by 0 days . dendritic cell transmigration ” , J Immunol ., Mar . 1 , 2014 , pp . 2366 2373, 192 ( 5 ) , The American Association of Immunologists , Inc ., (21 ) Appl. No. : 15 / 199 ,476 Rockville, MD . Pachynski et al. , “ The chemoattractant chemerin suppresses mela (22 ) Filed : Jun . 30 , 2016 noma by recruiting natural killer cell antitumor defenses” , The Journal of experimental medicine , Jul. 2 , 2012 , pp . 1427 - 1435 , (65 ) Prior Publication Data 209 ( 8 ) , The Rockefeller University Press, New York , NY. Del Prete et al. , “ CCRL2 , a fringe member of the atypical chemoat US 2017 / 0002087 A1 Jan . 5 , 2017 tractant receptor family ” , European Journal of immunology, Jun . 2013, pp . 1418 - 1422 , 43 ( 6 ), John Wiley & Sons , Inc ., Hoboken , NJ. Mattern et al ., “ Processing, signaling , and physiological function of Related U . S . Application Data chemerin ” , IUBMB Life , Jan . 20 , 2014 , pp . 19 - 26 , 66 ( 1 ) , John Wiley & Sons, Inc. , Hoboken , NJ. (60 ) Provisional application No . 62/ 186 , 734 , filed on Jun . Zabel et al . , “ Chemerin regulation and role in host defense ” , Am J 30 , 2015 . Clin Exp Immunol. , Mar . 15 , 2014 , pp . 1 - 19 , 3 ( 1 ) , e -Century Publishing Corporation , Madison , WI. (51 ) Int. CI. Monnier et al, “ Expression , regulation , and function of atypical COZK 16 / 30 ( 2006 .01 ) chemerin receptor CCRL2 on endothelial cells ” , Journal of immu A61K 39 /395 ( 2006 . 01 ) nology , Jul . 15 , 2012 , pp . 956 - 967, 189 ( 2 ) , The American Associa A61K 45 /06 ( 2006 . 01 ) tion of Immunologists , Inc ., Rockville , MD . A61K 9 / 00 ( 2006 .01 ) A61K 39 /00 ( 2006 .01 ) * cited by examiner (52 ) U . S . CI. CPC .. .. .. .. COZK 16 /3053 (2013 .01 ) ; A61K 9 / 00 Primary Examiner - Robert Landsman (2013 .01 ) ; A61K 39 / 39558 ( 2013 .01 ) ; A61K (74 ) Attorney , Agent, or Firm — Pamela J . Sherwood ; 45/ 06 ( 2013 .01 ) ; A61K 2039/ 505 (2013 . 01 ) ; Bozicevic , Field & Francis LLP A61K 2039/ 507 ( 2013 . 01 ) (58 ) Field of Classification Search (57 ) ABSTRACT CPC .. .. CO7K 16 /3053 ; A61K 39 /39558 ; A61K Chemerin is a chemoattractant that selectively directs intra 45 /06 : A61K 2039 / 505 ; A61K 2039 / 507 tumor recruitment of CMKLR1+ cells . Blockade of CCRL2 See application file for complete search history . from binding to chemerin in non -neoplastic tumor cells , e . g . peri - tumor stromal and / or hematopoietic cells allows for (56 ) References Cited redistribution of chemerin into the tumor and enhanced responsiveness against a tumor . FOREIGN PATENT DOCUMENTS 6 Claims, 7 Drawing Sheets WO WO2005005720 * 6 / 2005 ( 7 of 7 Drawing Sheet( s ) Filed in Color ) U .S . Patentatent Jan . 16 , 2018 Sheet 1 of 7 US 9 , 868 ,792 B2 CCRL2 Cancer Noma Analysis Type by Cancer warenestivererererererererererererererererererererer Bladder Cancer * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * rest vacy drevene Colorectal Cancer sexe StatistinitivamentarisatisiminiministrativniEsophagealCancer sistemin Gästric cances W Meno ano Neck Cancer dney Cancer Leukemia e UW Cancer event sanom Ovarian Cancer Paix Cance prosti Cancer Sarcomm Fig 1 U .S . Pateratent Jan . 16 , 2018 Sheet 2 of 7 US 9 , 868 , 792 B2 2001 WT r ion CCRL2 KO ment tumorsize(mma) Apertu ** * 3 10 13 16 days after tumor challenge m * * s WT CCRL240 Fig 2 U . S . Patent Jan . 16 , 2018 Sheet 3 of 7 US 9 ,868 ,792 B2 edforderland 2045%ETRL don www.ban CCRUZKO B WT CCRL24 Gated on C0454 01103 WUMWIKIMET F4 /80 Gated on CD11b * F4 /80 . * . C0119 . CO11b 0 wwws alsvous SXO A v vvvvvvvvvvvvvvvvvvvvvvvvvvvvvvvvvvvvvvv wwwwwww CD3 Fig 3 atent Jan . 16 , 2018 Sheet 4 of 7 US 9 , 868, 792 B2 pok W7 CCRL2 KO 10 * B Wiid To ?.* CCRUZKO ?? CD208 * ????????? ?????? 2 O 20 408 o 2012 *12 * - + - + - + + + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + * + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + * + - + + - + + - + - + - + - + - + - + - + * * * * * * * * * * * * * * MHC } Fig 4 atent Jan . 16 , 2018 Sheet 5 of 7 US 9 , 868, 792 B2 DAMC031DAPI 10X 20X WT Jou/sjassen * : : : :: : : :: : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : :: : : :: :: : :: : : :: : :: XOL 20X : : : : : : :: :: : : : .: : : : : : : : :: : 2 11111111111111 CCRL2KO . CCRL24 Fig 5 atent Jan . 16 , 2018 Sheet 6 of 7 US 9 , 868, 792 B2 Plasma B Tumor WTCCRL21 WT. CCRL2 Mouse # 123456 123456 p -value 123456 , 123456 D - value EDTAXIK YEGE 0 . 1 INIO CCL ? * 117 CCL2 w ** * * (Cytokine ) [ Cytokine ] www . ** pgm MCP3 p????? TGFE 1123 IL6 1122 LIX LIF HHHIL10 - -- - - 0. 08 IL22 114 -. * ** * 22 IL28 1ll2360* MI? - - - * ** * ILIB * w * CCL 1118 ILI8 W5F * CCLII WEGE 0 . 07 CCLS CSCF IFKG * 384 STUFA MCSF IL23 118 IL3 116 IFNA SONUS 0 .07 ILI TGED CMCSF GIL IL31 PAVIES & CXCL ) 5 CC14 TINC CXC1: 2 GUCSF GSCK ILA 112870 IL10 112 IL2Et . W ! ! TNN 1131 MISIE IPIO MIPIA IL15 118 I13 IL17) IL17 ) 1128 IL13 PTID ILIZA Plasma WT D 2572, 54 TL Tumor Homogenate * * * * * * CCRL2 - -- ?io} * * * { T : Relativeinduction Re{a} : : immmmmmmmmmmmmmmmm : : : .2 : 3 . 3 11011010101010101 -SF L Xu COLI L-19 ** MA TCFCCL7 CCLX VEGEN 1 - 18 Fig 6 atent Jan . 16 , 2018 Sheet 7 of 7 US 9 , 868 , 792 B2 ~ 39? ? ????28 3 % Two tissus CCR2- 03 WONS 303 pauoste 103 MW Slooo 195 Ana CMKIKI eudocus M - a8sexiated over ? ??? M e chamenti wwwwwwwwwwwwwwwwwwwwwwww w wwwwwww deficient _ 02 : cioèsduoso MYS :0 Fig 7 US 9 , 868 , 792 B2 METHODS OF ENHANCING ANTI - TUMOR coupled receptor chemokine receptor- like 1 (CMKLR1 ) , IMMUNITY BY ADMINISTERING which is expressed on several immune cells , in particular ANTIBODIES TO THE CCRL2 CHEMERIN NK cells . RECEPTOR A second chemerin -binding receptor, CC - chemokine 5 receptor - like 2 (CCRL2 ) is primarily expressed on endothe CROSS REFERENCE lial cells , activated macrophages, and mast cells . Unlike CMKLR1, CCRL2 does not internalize or trigger intracel This application claims benefit of U . S . Provisional Patent lular calcium mobilization upon binding chemerin . Instead , Application No . 62 / 186 ,734 . filed Jun . 30 . 2015 . now CCRL2 binds chemerin with high affinity and presents it on expired , this application is incorporated herein by reference 10 the cell surface . The present invention provides for a role of in its entirety . CCRL2 in cancer progression . GOVERNMENT RIGHTS SUMMARY 15 Methods are provided for immune targeting of cancer by This invention was made with Government support under contacting an individual with an agent that blocks CCRL2 , contract W81XWH - 11 - 1 - 0512 awarded by the Department and prevents CCRL2 from sequestering chemerin . It is of Defense and under contracts A1079320 and CA169354 shown herein that CCRL2 expression by the non -neoplastic , awarded by the National Institutes of Health . The Govern stromal cells associated with a tumor reduces anti -cancer ment has certain rights in the invention . 20 immune responses . The non - neoplastic cells may include peri - tumor fibroblasts and other connective tissue cells , BACKGROUND hematopoietic cells, etc . Blocking CCRL2 activity increases the bioavailability
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