A Rationally Designed Agonist Antibody Fragment That Functionally Mimics Thrombopoietin

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2006 Sep 16

PMID 16973749

Citations 14

Authors

Shana Frederickson

Mark W Renshaw

Bing Lin

Lynette M Smith

Peter Calveley

Jeremy P Springhorn

Krista Johnson

Yi Wang

Xiao Su

Yamin Shen

Katherine S Bowdish

Affiliations

Soon will be listed here.

Abstract

By using rational design, antibody fragments (Fabs) that mimic thrombopoietin (TPO) were created. A peptide with cMpl receptor-binding capability was grafted into different complementarity-determining regions of a fully human Fab scaffold. Functional presentation of the peptide was optimized by using phage display and cell-based panning. Select antibodies and fragments containing two grafted peptides were assayed for their ability to stimulate the cMpl receptor in vitro. Several candidates demonstrated agonist activity in an in vitro cMpl receptor signaling reporter assay, including Fab59, which was estimated to be equipotent to TPO. Fab59 additionally was able to effectively stimulate platelet production in normal mice. These rationally designed mimetic Fabs may provide a therapeutic intervention for thrombocytopenia while avoiding the potential generation of neutralizing antibodies to endogenous TPO. Furthermore, this study demonstrates a method by which short-lived linear peptides with binding activity may be converted to more stable and potent agonists capable of activating cell surface receptors.

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References

Verma I, Deschamps J, Van Beveren C, Sassone-Corsi P . Human fos gene. Cold Spring Harb Symp Quant Biol. 1986; 51 Pt 2:949-58. DOI: 10.1101/sqb.1986.051.01.108. View

Vadhan-Raj S, Verschraegen C, Bueso-Ramos C, Broxmeyer H, Kudelka A, Freedman R . Recombinant human thrombopoietin attenuates carboplatin-induced severe thrombocytopenia and the need for platelet transfusions in patients with gynecologic cancer. Ann Intern Med. 2000; 132(5):364-8. DOI: 10.7326/0003-4819-132-5-200003070-00005. View

Li J, Yang C, Xia Y, Bertino A, Glaspy J, Roberts M . Thrombocytopenia caused by the development of antibodies to thrombopoietin. Blood. 2001; 98(12):3241-8. DOI: 10.1182/blood.v98.12.3241. View

Barbas 3rd C, Rosenblum J, Lerner R . Direct selection of antibodies that coordinate metals from semisynthetic combinatorial libraries. Proc Natl Acad Sci U S A. 1993; 90(14):6385-9. PMC: 46936. DOI: 10.1073/pnas.90.14.6385. View

Feese M, Tamada T, Kato Y, Maeda Y, Hirose M, Matsukura Y . Structure of the receptor-binding domain of human thrombopoietin determined by complexation with a neutralizing antibody fragment. Proc Natl Acad Sci U S A. 2004; 101(7):1816-21. PMC: 357010. DOI: 10.1073/pnas.0308530100. View

McCullough J . Current issues with platelet transfusion in patients with cancer. Semin Hematol. 2000; 37(2 Suppl 4):3-10. DOI: 10.1016/s0037-1963(00)90047-7. View

Kuter D, Begley C . Recombinant human thrombopoietin: basic biology and evaluation of clinical studies. Blood. 2002; 100(10):3457-69. DOI: 10.1182/blood.V100.10.3457. View

Kimura T, Kaburaki H, Miyamoto S, Katayama J, Watanabe Y . Discovery of a novel thrombopoietin mimic agonist peptide. J Biochem. 1998; 122(5):1046-51. DOI: 10.1093/oxfordjournals.jbchem.a021845. View

de Serres M, Ellis B, Dillberger J, Rudolph S, Hutchins J, Boytos C . Immunogenicity of thrombopoietin mimetic peptide GW395058 in BALB/c mice and New Zealand white rabbits: evaluation of the potential for thrombopoietin neutralizing antibody production in man. Stem Cells. 1999; 17(4):203-9. DOI: 10.1002/stem.170203. View

10.

Wrighton N, Farrell F, Chang R, Kashyap A, Barbone F, Mulcahy L . Small peptides as potent mimetics of the protein hormone erythropoietin. Science. 1996; 273(5274):458-64. DOI: 10.1126/science.273.5274.458. View

11.

Wilson I, Niman H, Houghten R, Cherenson A, Connolly M, Lerner R . The structure of an antigenic determinant in a protein. Cell. 1984; 37(3):767-78. DOI: 10.1016/0092-8674(84)90412-4. View

12.

Erickson-Miller C, Delorme E, Tian S, Hopson C, Stark K, Giampa L . Discovery and characterization of a selective, nonpeptidyl thrombopoietin receptor agonist. Exp Hematol. 2005; 33(1):85-93. DOI: 10.1016/j.exphem.2004.09.006. View

13.

Wang B, Nichol J, Sullivan J . Pharmacodynamics and pharmacokinetics of AMG 531, a novel thrombopoietin receptor ligand. Clin Pharmacol Ther. 2004; 76(6):628-38. DOI: 10.1016/j.clpt.2004.08.010. View

14.

Dorsch M, Fan P, Danial N, Rothman P, Goff S . The thrombopoietin receptor can mediate proliferation without activation of the Jak-STAT pathway. J Exp Med. 1998; 186(12):1947-55. PMC: 2199166. DOI: 10.1084/jem.186.12.1947. View

15.

Barbas 3rd C, Languino L, Smith J . High-affinity self-reactive human antibodies by design and selection: targeting the integrin ligand binding site. Proc Natl Acad Sci U S A. 1993; 90(21):10003-7. PMC: 47701. DOI: 10.1073/pnas.90.21.10003. View

16.

Evans A, Krentz A . Recent developments and emerging therapies for type 2 diabetes mellitus. Drugs R D. 2000; 2(2):75-94. DOI: 10.2165/00126839-199902020-00001. View

17.

Broudy V, Lin N . AMG531 stimulates megakaryopoiesis in vitro by binding to Mpl. Cytokine. 2003; 25(2):52-60. DOI: 10.1016/j.cyto.2003.05.001. View

18.

Scheding S, Mohle R, Buhring H, Baum C, Mc Kearn J, Buchner T . Effective ex vivo generation of megakaryocytic cells from mobilized peripheral blood CD34(+) cells with stem cell factor and promegapoietin. Exp Hematol. 2000; 28(3):335-46. DOI: 10.1016/s0301-472x(99)00152-6. View

19.

Begley C, Basser R . Biologic and structural differences of thrombopoietic growth factors. Semin Hematol. 2000; 37(2 Suppl 4):19-27. DOI: 10.1016/s0037-1963(00)90049-0. View

20.

Vadhan-Raj S, Murray L, Bueso-Ramos C, Patel S, Reddy S, Hoots W . Stimulation of megakaryocyte and platelet production by a single dose of recombinant human thrombopoietin in patients with cancer. Ann Intern Med. 1997; 126(9):673-81. DOI: 10.7326/0003-4819-126-9-199705010-00001. View