Methods and Compositions for Inhibiting Cell Death or Enhacing Cell Proliferation
The present invention provides compositions and methods that enhance cell survival. Such compositions feature chimeric polypeptides that include at least a GM-CSF receptor ligand and an anti-apoptotic moiety (e.g., a Bcl-2 protein family member). In one embodiment, the chimeric polypeptide is a GM-CSF-Bcl-xL chimeric polypeptide. The invention further includes methods of using chimeric polypeptides to enhance cell survival or inhibit cell death in a cell at risk of cell death.
1 . An isolated chimeric polypeptide comprising a GM-CSF receptor ligand or polypeptide and a Bcl-xL polypeptide, wherein the chimeric polypeptide specifically binds a GM-CSF receptor and enhances cell survival.
2 - 7 . (canceled)
8 . The isolated chimeric polypeptide of claim 1 , wherein the chimeric polypeptide inhibits cell death.
9 - 12 . (canceled)
13 . The chimeric polypeptide of claim 1 , wherein the polypeptide comprises at least a fragment of Bcl-xL capable of inhibiting cell death.
14 - 16 . (canceled)
17 . The chimeric polypeptide of claim 1 , wherein the polypeptide comprises a fragment selected from the group consisting of
i.
(SEQ ID NO: 19)
APARSPSPSTQPWEHVNAIQEARRLLNLSRDTAAEMNETVEVISEMFD
LQEPTCLQTRLELYKQGLRGSLTKLKGPLTMMASHYKQHCPPTPETSC
ATQTITFESFKENLKDFLLVIPFDCWEPVQE;
ii.
(SEQ ID NO: 20)
EARRLLNLSRD;
and
iii.
(SEQ ID NO: 4)
TMMASHYKQHCPPTPET.
18 . The chimeric polypeptide of claim 1 , wherein the polypeptide consists essentially of an active fragment of GM-CSF selected from the group consisting of:
i.
(SEQ ID NO: 19)
APARSPSPSTQPWEHVNAIQEARRLLNLSRDTAAEMNETVEVISEMFD
LQEPTCLQTRLELYKQGLRGSLTKLKGPLTMMASHYKQHCPPTPETSC
ATQTITFESFKENLKDFLLVIPFDCWEPVQE;
ii.
(SEQ ID NO: 20)
EARRLLNLSRD;
and
iii.
(SEQ ID NO: 4)
TMMASHYKQHCPPTPET.
19 - 30 . (canceled)
31 . An isolated nucleic acid molecule that encodes a chimeric polypeptide of claim 1 .
32 - 37 . (canceled)
37 . The isolated nucleic acid molecule of claim 36 , comprising a nucleic acid molecule having substantial nucleic acid sequence identity to SEQ ID NO: 10.
38 - 40 . (canceled)
41 . An isolated polynucleotide capable of encoding a polypeptide having substantial sequence identity to SEQ ID NO: 1, wherein the polypeptide enhances cell survival, promotes cell proliferation, or inhibits cell death.
42 . A vector comprising a nucleic acid molecule that encodes a polypeptide of claim 1 .
43 - 47 . (canceled)
48 . A host cell comprising the vector of claim 42 .
49 - 55 . (canceled)
56 . A pharmaceutical composition comprising an effective amount of a chimeric polypeptide of claim 1 , or fragments thereof, in a pharmaceutically acceptable excipient.
57 . A pharmaceutical composition comprising an effective amount of a nucleic acid molecule encoding a chimeric polypeptide of claim 1 in a pharmaceutically acceptable excipient.
58 . The pharmaceutical composition of claim 56 , wherein the composition further comprises an agent selected from the group consisting of a chemotherapeutic agent, radiation agent, hormonal agent, biological agent, an anti-inflammatory agent, an agent that enhances dopamine production, an anticholinergic, a dopamine mimetic, amantadine, an antithrombotic, and a thrombolytic.
59 . A method of enhancing cell survival, the method comprising contacting a cell at risk of cell death with a chimeric polypeptide of claim 1 , wherein the contacting enhances cell survival.
60 . A method of inhibiting cell death in a cell at risk thereof, the method comprising contacting the cell at risk of cell death with a chimeric polypeptide of claim 1 , wherein the contacting inhibits cell death.
61 . A method of enhancing cell survival, the method comprising contacting a cell at risk of cell death with a nucleic acid molecule of claim 28 , wherein the contacting enhances cell survival.
62 . A method of inhibiting cell death in a cell at risk thereof, the method comprising contacting the cell with a nucleic acid molecule of claim 28 , wherein the contacting inhibits cell death.
63 - 69 . (canceled)
70 . A method of enhancing cell survival in a subject diagnosed as having a disease or disorder characterized by cell death, the method comprising administering to the subject a chimeric polypeptide of claim 1 in an amount effective to enhance cell survival.
71 . A method of enhancing cell survival in a subject diagnosed as having a disease or disorder characterized by cell death, the method comprising administering to the subject a nucleic acid molecule encoding the chimeric polypeptide of claim 1 in an amount effective to enhance cell survival.
72 - 75 . (canceled)
76 . A method of assessing the efficacy of a cell survival enhancing treatment in a subject, comprising:
determining one or more pre-treatment phenotypes;
administering a therapeutically effective amount of a chimeric polypeptide of claim 1 , or a nucleic acid molecule encoding the polypeptide to the subject; and
determining the one or more phenotypes after an initial period of treatment with the an cell death inhibitor; wherein the modulation of the one or more phenotypes indicates efficacy of a cell death inhibitor treatment.
77 . A method of selecting a subject having a disease or disorder characterized by cell death for treatment with a cell death inhibitor, comprising:
determining one or more pre-treatment phenotypes,
administering a therapeutically effective amount of a chimeric polypeptide of claim 1 , or a nucleic acid molecule encoding the polypeptide to the subject; and
determining the one or more phenotypes after an initial period of treatment with the cell death inhibitor, wherein the modulation of the one or more phenotype is an indication that the disorder is likely to have a favorable clinical response to treatment with a cell death inhibitor.
78 - 91 . (canceled)
92 . A method of expanding hematopoietic stem cells or progenitor cells comprising contacting the cells with an effective amount of a polypeptide or nucleic acid molecule of claim 1 .