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-12098179-B2
Application number	US-201916981474-A
Country	US
Kind code	B2
Filing date	Mar 15, 2019
Priority date	Mar 16, 2018
Publication date	Sep 24, 2024
Grant date	Sep 24, 2024

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

The invention claimed is:

1. A fusion protein comprising a peptide fused to a heterologous polypeptide or an immunoconjugate comprising an antibody fused or conjugated to the peptide, wherein the peptide comprises at least 8 consecutive amino acids of urocortin 3 (UCN3), and wherein:
the at least 8 consecutive amino acids comprise 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
the at least 8 consecutive amino acids comprise 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
the at least 8 consecutive amino acids comprise 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);
and wherein the peptide is not SEQ ID NO: 2 (MLMPVHFL), SEQ ID NO: 3 (MLMPVHFLL) or SEQ ID NO: 7 (LMPVHFLL).

2. The fusion protein or the immunoconjugate of claim 1 , wherein the peptide consists of the amino acid sequence as set forth in SEQ ID NO: 4 (MPVHFLLL) or SEQ ID NO: 34 (SLLSKRSFHY).

3. The fusion protein or the immunoconjugate of claim 1 , wherein the peptide consists of the amino acid sequence as set forth in SEQ ID NO: 4 (MLMPVHFLLL), SEQ ID NO: 5 (MLMPVHFLLLL), SEQ ID NO: 6 (FLLLLLLLL), 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: 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: 66 (LSEAEKGQWEDASL), SEQ ID NO: 67 (SRDASSGEEEEGKEKKTFPISGARGGARGTRYRYVSQAQPRGKPRQDTAKSPHR TK), 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. The immunoconjugate of claim 1 , wherein the antibody is directed against a surface antigen of an antigen presenting cell so that the peptide is targeted to said antigen presenting cell to elicit an immune response.

5. A pharmaceutical or vaccine composition comprising the fusion protein or the immunoconjugate of claim 1 .

6. The immunoconjugate of claim 4 , wherein the immune response is tolerance.

7. A composition comprising a peptide and an adjuvant, wherein the peptide comprises at least 8 consecutive amino acids of urocortin 3 (UCN3), and wherein:
the at least 8 consecutive amino acids comprise 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
the at least 8 consecutive amino acids comprise 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
the at least 8 consecutive amino acids comprise 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);
and wherein the peptide is not SEQ ID NO: 2 (MLMPVHFL), SEQ ID NO: 3(MLMPVHFLL) or SEQ ID NO: 7 (LMPVHFLL).

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

• C12N2310/16

  Aptamers · 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

• C07K16/26

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

• C07K14/70539

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

• 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