Polypeptides and uses thereof for reducing CD95-mediated cell motility

Patent metadata

Field	Value
Publication number	US-10189887-B2
Application number	US-201515304550-A
Country	US
Kind code	B2
Filing date	Apr 16, 2015
Priority date	Apr 17, 2014
Publication date	Jan 29, 2019
Grant date	Jan 29, 2019

Abstract

The present invention relates to polypeptides and uses thereof for reducing CD95-meditated cell motility. In particular, the present invention relates to a polypeptide having an amino acid sequence having at least 70% of identity with the amino acid sequence ranging from the amino-acid residue at position 175 to the amino-acid residue at position 191 in SEQ ID NO:1.

First claim

The invention claimed is:

1. A fusion protein, comprising a polypeptide having an amino acid sequence having at least 96, 97, 98, 99 or 100% identity with an amino acid sequence ranging from an amino-acid residue at position 175 to an amino-acid residue at position 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, or 210 as set forth in SEQ ID NO:1,
wherein said polypeptide is fused to a heterologous cell-penetrating peptide.

2. The fusion protein of claim 1 , wherein said polypeptide has the amino acid identity of amino-acid residues 175 to 191 as set forth in SEQ ID NO: 1.

3. The fusion protein of claim 1 , wherein said polypeptide has the amino acid identity of amino-acid residues 175 to 210 as set forth in SEQ ID NO: 1.

4. The fusion protein of claim 3 , having said amino-acid residues 175 to 210 fused to said heterologous cell-penetrating peptide, wherein said heterologous cell-penetrating peptide is TAT, having a sequence identity as set forth in SEQ ID NO: 3 or SEQ ID NO:4.

5. The fusion protein of claim 1 , wherein said heterologous cell-penetrating peptide is selected from the group consisting of penetratin; TAT mitochondrial penetrating sequence; an internalization sequence derived from Drosophila Antennapedia/Penetratin (Antp) protein, having a sequence identity as set forth in SEQ ID NO: 2; and Transactivator of Transcription (TAT), having a sequence identity as set forth in SEQ ID NO:3 or SEQ ID NO: 4.

6. The fusion protein of claim 1 , wherein said cell-penetrating peptide is TAT, having a sequence identity as set forth in SEQ ID NO: 3 or SEQ ID NO: 4.

7. A fusion protein comprising a polypeptide having an amino acid sequence having at least 96, 97, 98, 99 or 100% identity with an amino acid sequence ranging from an amino-acid residue at position 175 to an amino-acid residue at position 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, or 210 as set forth in SEQ ID NO:1 fused directly or via a spacer to at least one heterologous polypeptide.

8. The fusion protein of claim 7 wherein the at least one heterologous polypeptide is a cell-penetrating peptide, a Transactivator of Transcription (TAT) cell penetrating sequence, a cell permeable peptide or a membranous penetrating sequence.

9. The fusion protein of claim 8 , wherein said heterologous peptide is TAT, having a sequence identity as set forth in SEQ ID NO: 3 or SEQ ID NO: 4.

10. A TAT-CID fusion protein, having a sequence identity as set forth in SEQ ID NO: 6.

11. A method for producing a fusion protein comprising
a polypeptide having an amino acid sequence ranging from an amino-acid residue at position 175 to an amino-acid residue at position 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210 as set forth in SEQ ID NO:1, and
a cell-penetrating peptide,

the method comprising the steps of:
(i) culturing a transformed host cell comprising a nucleic acid sequence encoding the fusion protein comprising the polypeptide and the cell-penetrating peptide, or a vector comprising an expression cassette comprising the nucleic acid sequence, under conditions suitable to allow expression of said fusion protein; and
(ii) recovering the expressed fusion protein.

Assignees

• Inst Nat Sante Rech Med
• Univ Bordeaux
• Univ Rennes
• Ecole Des Hautes Etudes En Sante Publique Ehesp
• Univ Des Antilles Et De La Guyane
• Inst Bergonie
• Univ Nottingham
• Inst Nat Sante Rech Med
• Univ Bordeaux
• Univ Rennes
• Ecole Des Hautes Etudes En Sante Publique Ehesp
• Univ Des Antilles Et De La Guyane
• Inst Bergonie

Inventors

• Legembre Patrick
• Vacher Pierre
• Sanseau Doriane
• Penna Aubin
• Flynn Robin

Classifications

• G01N2500/02

  Screening involving studying the effect of compounds C on the interaction between interacting molecules A and B (e.g. A = enzyme and B = substrate for A, or A = receptor and B = ligand for the receptor) · CPC title

• G01N2333/70575

  NGF/TNF-superfamily, e.g. CD70, CD95L, CD153 or CD154 · CPC title

• G01N33/6872

  Intracellular protein regulatory factors and their receptors, e.g. including ion channels · CPC title

• C07K14/525 Primary

  Tumour necrosis factor [TNF] · CPC title

• C07K14/70575 Primary

  NGF/TNF-superfamily, e.g. CD70, CD95L, CD153, CD154 (NGF C07K14/48, TNF C07K14/525) · CPC title