Antigenic peptides deriving from urocortin 3 and uses thereof for the diagnosis and treatment of type 1 diabetes

Patent metadata

Field	Value
Publication number	US-2021024603-A1
Application number	US-201916981474-A
Country	US
Kind code	A1
Filing date	Mar 15, 2019
Priority date	Mar 16, 2018
Publication date	Jan 28, 2021
Grant date	—

Abstract

Despite the notion that human CD8+ T cells are the final mediators of autoimmune β-cell destruction in type 1 diabetes (T1D), none of their target epitopes has been demonstrated to be naturally processed and presented by β cells. The inventors therefore performed an epitope discovery study combining HLA Class I peptidomics and transcriptomics strategies. Inflammatory cytokines increased β-cell peptide presentation in vitro, paralleling upregulation of HLA Class I expression. Peptide sources included known β-cell antigens and several insulin granule proteins. Urocortin 3 was identified as a novel β-cell antigen, which was processed into HLA-A2- and HLA-A3-restricted epitopes recognized by circulating naive CD8+ T cells in type 1 diabetic and healthy donors. Accordingly, the present invention relates to antigenic peptides derived from urocortin-3 and uses thereof for the diagnosis and treatment of T1D.

First claim

1 . A peptide derived from urocortin 3 (UCN3) comprising:
at least 8 consecutive amino acids in the sequence ranging from the amino acid residue at position 1 to the amino acid residue at position 21 in SEQ ID NO:1 (UCN3), or at least 8 consecutive amino acids in the sequence ranging from the amino acid residue at position 22 to the amino acid residue at position 71 in SEQ ID NO:1 (UCN3), or at least 8 consecutive amino acids in the sequence ranging from the amino acid residue at position 119 to the amino acid residue at position 162 in SEQ ID NO:1 (UCN3).

2 . The isolated peptide of claim 1 which consists of the amino acid sequence as set forth in SEQ ID NO: 4 (MLMPVHFLLL) or SEQ ID NO: 34 (SLLSKRSFHY).

3 . The isolated peptide of claim 1 which consists of the amino acid sequence as set forth in SEQ ID NO: 2 (MLMPVHFL), SEQ ID NO: 3 (MLMPVHFLL), SEQ ID NO: 4 (MLMPVHFLLL), SEQ ID NO: 5 (MLMPVHFLLLL), SEQ ID NO: 6 (FLLLLLLLL), SEQ ID NO: 7 (LMPVHFLL), SEQ ID NO: 8 (LMPVHFLLL), SEQ ID NO: 9 (LMPVHFLLLL), SEQ ID NO: 10 (HFLLLLLLLL), SEQ ID NO: 11 (FLLLLLLL), SEQ ID NO: 12 (FLLLLLLLLG), SEQ ID NO: 13 (LLLGGPRTGL), SEQ ID NO: 14 (PRTGLPHKFYK), SEQ ID NO: 15 (RTGLPHKFYK), SEQ ID NO: 16 (GLPHKFYKAK), SEQ ID NO: 17 (MLMPVHFL), SEQ ID NO: 18 (MLMPVHFLL), SEQ ID NO: 19 (MLMPVHFLLL), SEQ ID NO: 20 (MPVHFLLL), SEQ ID NO: 21 (FLLLLLLLLGGPRTG), SEQ ID NO: 22 (LLLLLLLLGGPRTGL), SEQ ID NO: 23 (LLLLLLLGGPRTGLP), SEQ ID NO: 24 (LLLLLLGGPRTGLPH), SEQ ID NO: 25 (GLPHKFYKAKPIFSC), SEQ ID NO: 26 (LPHKFYKAKPIFSCL), SEQ ID NO: 27 (GLPHKFYKAKPIFSC), SEQ ID NO: 28 (LPHKFYKAKPIFSCL), SEQ ID NO: 29 (PRTGLPHKFY), SEQ ID NO: 30 (GQWEDASLL), SEQ ID NO: 31 (SLLSKRSFHYL), SEQ ID NO: 32 (LLSKRSFHYL), SEQ ID NO: 33 (GQWEDASLLSK), SEQ ID NO: 34 (SLLSKRSFHY), SEQ ID NO: 35 (LLSKRSFHY), SEQ ID NO: 36 (RSFHYLRSR), SEQ ID NO: 37 (KFYKAKPIF), SEQ ID NO: 38 (YKAKPIFSCL), SEQ ID NO: 39 (SLLSKRSF), SEQ ID NO: 40 (LLSKRSFHYL), SEQ ID NO: 41 (YLRSRDASS), SEQ ID NO: 42 (PHKFYKAKPIFSCLN), SEQ ID NO: 43 (HKFYKAKPIFSCLNT), SEQ ID NO: 44 (KFYKAKPIFSCLNTA), SEQ ID NO: 45 (LSKRSFHYLRSRDAS), SEQ ID NO: 46 (SKRSFHYLRSRDASS), SEQ ID NO: 47 (KRSFHYLRSRDASSG), SEQ ID NO: 48 (RSFHYLRSRDASSGE), SEQ ID NO: 49 (SFHYLRSRDASSGEE), SEQ ID NO: 50 (EDASLLSKRSFHYLR), SEQ ID NO: 51 (DASLLSKRSFHYLRS), SEQ ID NO: 52 (ASLLSKRSFHYLRSR), SEQ ID NO: 53 (PHKFYKAKPIFSCLN), SEQ ID NO: 54 (HKFYKAKPIFSCLNT), SEQ ID NO: 55 (YKAKPIFSCLNTALS), SEQ ID NO: 56 (KAKPIFSCLNTALSE), SEQ ID NO: 57 (AKPIFSCLNTALSEA), SEQ ID NO: 58 (KPIFSCLNTALSEAE), SEQ ID NO: 59 (PIFSCLNTALSEAEK), SEQ ID NO: 60 (LSKRSFHYLRSRDAS), SEQ ID NO: 61 (SKRSFHYLRSRDASS), SEQ ID NO: 62 (KRSFHYLRSRDASSG), SEQ ID NO: 63 (RSFHYLRSRDASSGE), SEQ ID NO: 64 (SFHYLRSRDASSGEE), SEQ ID NO: 65 (PRTGLPHKFY), SEQ ID NO: (LSEAEKGQWEDASL), SEQ ID NO: 67 (SRDAS S GEEEEGKEKKTFPIS GARGGARGTRYRYVS QAQPRGKPRQDTA KSPHRTK), SEQ ID NO: 68 (TLSLDVPTNI), SEQ ID NO: 69 (TNIMNLLFNI), SEQ ID NO: 70 (NIMNLLFNI), SEQ ID NO: 71 (IMNLLFNI), SEQ ID NO: 72 (IMNLLFNIAK), SEQ ID NO: 73 (MNLLFNIAKAK), SEQ ID NO: 74 (NLLFNIAKAK), SEQ ID NO: 75 (LLFNIAKAK), SEQ ID NO: 76 (AHLMAQIGRK), SEQ ID NO: 77 (HLMAQIGRK), SEQ ID NO: 78 (HLMAQIGRKK), SEQ ID NO: 79 (LMAQIGRKK), SEQ ID NO: 80 (IMNLLFNIAKAKNLR), SEQ ID NO: 81 (MNLLFNIAKAKNLRA), SEQ ID NO: 82 (NLLFNIAKAKNLRAQ), SEQ ID NO: 83 (LLFNIAKAKNLRAQA), SEQ ID NO: 84 (LFNIAKAKNLRAQAA), SEQ ID NO: 85 (AKAKNLRAQAAANAH), SEQ ID NO: 86 (KAKNLRAQAAANAHL), SEQ ID NO: 87 (AKNLRAQAAANAHLM), SEQ ID NO: 88 (KNLRAQAAANAHLMA), SEQ ID NO: 89 (FTLSLDVPTNIMNLL), SEQ ID NO: 90 (TLSLDVPTNIMNLLF), SEQ ID NO: 91 (IMNLLFNIAKAKNLR), SEQ ID NO: 92 (MNLLFNIAKAKNLRA), SEQ ID NO: 93 (NLLFNIAKAKNLRAQ), SEQ ID NO: 94 (LLFNIAKAKNLRAQA), SEQ ID NO: 95 (LFNIAKAKNLRAQAA), SEQ ID NO: 96 (MNLLFNIAKAKNLRA), SEQ ID NO: 97 (NLLFNIAKAKNLRAQ), SEQ ID NO: 98 (LLFNIAKAKNLRAQA), SEQ ID NO: 99 (LFNIAKAKNLRAQAA), SEQ ID NO: 100 (KAKNLRAQAAANAHL), SEQ ID NO: 101 (AKNLRAQAAANAHLM), SEQ ID NO: 102 (KNLRAQAAANAHLMA), or SEQ ID NO: 103 (NLRAQAAA).

4 . A fusion protein comprising the peptide of claim 1 fused to a heterologous polypeptide.

5 . An immunoconjugate comprising an antibody fused or conjugated to the peptide of claim 1 .

6 . The immunoconjugate of claim 5 wherein the antibody is directed against a surface antigen of an antigen presenting cell so that the peptide of claim 1 is targeted to said antigen presenting cell to elicit an immune response (e.g. tolerance).

7 . An aptamer or an antibody having specificity for the peptide of claim 1 , either alone or complexed with HLA molecules that are permissive for peptide binding.

8 . A chimeric antigen receptor (CARs) comprising an antigen binding domain of the antibody of claim 7 .

9 . A T-cell receptor (TCR) having specificity for the peptide of claim 1 .

10 . A nucleic acid that encodes for the peptide of claim 1 , a fusion protein comprising the peptide, a chimeric antigen receptor comprising an antigen binding domain of an antibody having specificity for the peptide or a TCR having specificity for the peptide.

11 . A host cell comprising the nucleic acid of claim 10 , wherein the nucleic acid encodes the chimeric antigen receptor or the TCR.

12 . The host cell of claim 11 which is T cell.

13 . A MHC class I or class II multimer loaded with the peptide of claim 1 .

14 . A method of treating type 1 diabetes in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the peptide of claim 1 , a fusion protein comprising the peptide, an immunoconjugate comprising an antibody fused or conjugated to the peptide, a population of host cells comprising a nucleic acid encoding the peptide or a fusion protein comprising the peptide or a chimeric antigen receptor comprising an antigen binding domain of an antibody having specificity for the peptide or a TCR having specificity for the peptide, or an MHC class I or class II multimer loaded with the peptide.

15 . A pharmaceutical or vaccine composition comprising the peptide of claim 1 , a fusion protein comprising the peptide or an immunoconjugate comprising an antibody fused or conjugated to the peptide.

16 . (canceled)

17 . The host cell of claim 12 wherein the T cell is a Treg cell or a stem cell.

18 . A pharmaceutical or vaccine composition comprising a population of host cells of claim 11 .

19 . A pharmaceutical or vaccine composition comprising the MHC class I or class II multimer of claim 13 .

20 . The peptide of claim 1 , which is an isolated peptide.

Assignees

• Inst Nat Sante Rech Med
• Univ Paris
• Centre Nat Rech Scient
• Ecole Superieure Physique & Chimie Ind Ville De Paris

Inventors

• Mallone Roberto
• Gonzalez-Duque Sergio
• Verdier Yann
• Azoury Marie-Eliane
• Afonso Georgia
• Vinh Joëlle

Classifications

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title

• C07K2319/00

  Fusion polypeptide · CPC title

• C07K16/26

  against hormones {; against hormone releasing or inhibiting factors} · CPC title

• C12N15/115

  Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith {; Nucleic acids binding to non-nucleic acids, e.g. aptamers} · CPC title

• C07K14/70539

  MHC-molecules, e.g. HLA-molecules · CPC title