TREATMENT OF NEUROPATHY WITH IGF-1-ENCODING DNA CONSTRUCTS AND HGF-ENCODING DNA CONSTRUCTS

1 . A method of treating neuropathy, comprising the steps of:

administering to a subject having neuropathy a therapeutically effective amount of a first IGF-1-encoding DNA construct capable of expressing a human IGF-1 isoform; and

administering to the subject a therapeutically effective amount of first HGF-encoding DNA construct capable of expressing a human HGF isoform.

2 . The method of claim 1 , wherein the first IGF-1-encoding DNA construct is capable of expressing Class I IGF-1Ea protein comprising a polypeptide of SEQ ID NO: 14 or Class I IGF-1Ec protein comprising a polypeptide of SEQ ID NO: 16.

3 . (canceled)

4 . The method of claim 1 , wherein the first IGF-1-encoding DNA construct comprises a polynucleotide of SEQ ID NO: 15, optionally wherein the method further comprises the step of:

administering to the subject a second IGF-1-encoding DNA construct, wherein the second IGF-1-encoding DNA construct comprises a polynucleotide of SEQ ID NO: 17.

5 . (canceled)

6 . The method of claim 1 , wherein the first IGF-1-encoding DNA construct comprises a polynucleotide of SEQ ID NO: 17, optionally wherein the method further comprises the step of:

administering to the subject a second IGF-1-encoding DNA construct, wherein the second IGF-1-encoding DNA construct comprises a polynucleotide of SEQ ID NO: 15.

7 - 9 . (canceled)

10 . The method of claim 1 , wherein the first IGF-1-encoding DNA construct encodes more than one human IGF-1 isoforms comprising a polypeptide of SEQ ID NO: 14 and a polypeptide of SEQ ID NO: 16.

11 . (canceled)

12 . The method of claim 10 , wherein the first IGF-1-encoding DNA construct comprises:

a first IGF polynucleotide of SEQ ID NO: 1 (exons 1, 3, 4) or a degenerate thereof;

a second IGF polynucleotide of SEQ ID NO: 2 (intron 4) or a fragment thereof;

a third IGF polynucleotide of SEQ ID NO: 3 (exons 5 and 6-1) or a degenerate thereof;

a fourth IGF polynucleotide of SEQ ID NO: 4 (intron 5) or a fragment thereof; and

a fifth IGF polynucleotide of SEQ ID NO: 5 (exon 6-2) or a degenerate thereof,

wherein the first polynucleotide, the second polynucleotide, the third polynucleotide, the fourth polynucleotide and the fifth polynucleotide are linked in sequential 5′ to 3′ order.

13 . The method of claim 12 , wherein the second IGF polynucleotide is a polynucleotide of SEQ ID NO: 6, or SEQ ID NO: 7.

14 . (canceled)

15 . The method of claim 12 , wherein the fourth IGF polynucleotide is a polynucleotide of SEQ ID NO: 8.

16 - 18 . (canceled)

19 . The method of claim 12 , wherein the first IGF-1-encoding DNA construct comprises a polynucleotide of SEQ ID NO: 10 or SEQ ID NO: 9.

20 . (canceled)

21 . The method of claim 1 , wherein the first IGF-1-encoding DNA construct and the first HGF-encoding DNA construct are administered in an amount sufficient to reduce pain in the subject, optionally wherein the subject has diabetic neuropathy.

22 - 25 . (canceled)

26 . The method of claim 1 , wherein the first HGF-encoding DNA construct encodes two human HGF isoforms, wherein the two human HGF isoforms are flHGF of SEQ ID NO: 11 and dHGF of SEQ ID NO: 12.

27 . (canceled)

28 . The method of claim 1 , wherein the first HGF-encoding DNA construct comprises:

a first HGF polynucleotide of SEQ ID NO: 22 (exons 1-4) or a degenerate thereof;

a second HGF polynucleotide of SEQ ID NO: 25 (intron 4) or a functional fragment thereof; and

a third HGF polynucleotide of SEQ ID NO: 23 (exons 5-18) or a degenerate thereof,

wherein the second HGF polynucleotide is located between the first HGF polynucleotide and the third HGF polynucleotide, and the first HGF-encoding DNA construct encodes two human HGF isoforms.

29 . The method of claim 28 , wherein the first HGF-encoding DNA construct comprises a polynucleotide of SEQ ID NO: 13.

30 . The method of claim 1 , wherein the first IGF-1-encoding DNA construct and the first HGF-encoding DNA construct are co-administered, optionally via an intramuscular injection.

31 . (canceled)

32 . The method of claim 1 , wherein the step of administering the first IGF-1-encoding DNA construct and the step of administering the first HGF-encoding DNA construct are performed separately, optionally at least three weeks apart.

33 - 34 . (canceled)

35 . The method of claim 1 , comprising the steps of:

administering to a subject having neuropathy an HGF-encoding DNA construct comprising a polynucleotide of SEQ ID NO: 13 or a polynucleotide of SEQ ID NO: 33; and

administering to the subject an IGF-1-encoding DNA construct comprising a polynucleotide of SEQ ID NO: 10 or a polynucleotide of SEQ ID NO: 9,

wherein the step of administering the HGF-encoding DNA construct and the step of administering the IGF-1-encoding DNA construct are performed at least three weeks apart.

36 . (canceled)

37 . The method of claim 1 , comprising the steps of:

administering to a subject having neuropathy an HGF-encoding DNA construct comprising a polynucleotide of SEQ ID NO: 13; and

administering to the subject a first IGF-1-encoding DNA construct comprising a polynucleotide of SEQ ID NO: 15 and a second IGF-1-encoding DNA construct comprising a polynucleotide of SEQ ID NO: 17,

wherein the step of administering the HGF-encoding DNA construct and the step of administering the first IGF-1-encoding DNA construct and the second IGF-1-encoding DNA construct are performed at least three weeks apart.

38 . A pharmaceutical composition comprising:

an IGF-1-encoding DNA construct capable of expressing at least one human IGF-1 isoform;

an HGF-encoding DNA construct capable of expressing at least one human HGF isoform, and

a pharmaceutically acceptable excipient.

39 . The pharmaceutical composition of claim 38 , wherein the IGF-1-encoding DNA construct encodes Class I IGF-1Ea protein comprising a polypeptide of SEQ ID NO: 14, Class I IGF-1Ec protein comprising a polypeptide of SEQ ID NO: 16 or both Class I IGF-1Ea protein and Class I IGF-1Ec protein.

40 - 41 . (canceled)

42 . The pharmaceutical composition of claim 38 , wherein the IGF-1-encoding DNA construct, comprising:

a first IGF polynucleotide of SEQ ID NO: 1 (exons 1, 3, 4) or a degenerate thereof;

a second IGF polynucleotide of SEQ ID NO: 2 (intron 4) or a fragment thereof;

a third IGF polynucleotide of SEQ ID NO: 3 (exons 5 and 6-1) or a degenerate thereof;

a fourth IGF polynucleotide of SEQ ID NO: 4 (intron 5) or a fragment thereof; and

a fifth IGF polynucleotide of SEQ ID NO: 5 (exon 6-2) or a degenerate thereof,

wherein the first polynucleotide, the second polynucleotide, the third polynucleotide, the fourth polynucleotide and the fifth polynucleotide are linked in sequential 5′ to 3′ order.

43 . The pharmaceutical composition of claim 42 , wherein the second IGF polynucleotide is a polynucleotide of SEQ ID NO: 6, or SEQ ID NO: 7.

44 . (canceled)

45 . The pharmaceutical composition of claim 42 , wherein the fourth IGF polynucleotide is a polynucleotide of SEQ ID NO: 8.

46 - 47 . (canceled)

48 . The pharmaceutical composition of claim 38 , wherein the IGF-1-encoding DNA construct is selected from the group consisting of pCK-IGF-1X6, pCK-IGF-1X10, pTx-IGF-1X6 and pTx-IGF-1X10.

49 - 54 . (canceled)

55 . The pharmaceutical composition of claim 38 , wherein the HGF-encoding DNA construct comprises:

a first HGF polynucleotide of SEQ ID NO: 22 (exons 1-4) or a degenerate thereof;

a second HGF polynucleotide of SEQ ID NO: 25 (intron 4) or a functional fragment thereof; and

a third HGF polynucleotide of SEQ ID NO: 23 (exons 5-18) or a degenerate thereof,

wherein the second HGF polynucleotide is located between the first HGF polynucleotide and the third HGF polynucleotide, and the first HGF-encoding DNA construct encodes two human HGF isoforms.

56 . The pharmaceutical composition of claim 38 ,

wherein the HGF-encoding DNA construct comprises a polynucleotide of any of SEQ ID Nos: 26-32 and 13.

57 - 62 . (canceled)

63 . A kit for treating neuropathy, comprising:

a first pharmaceutical composition comprising an IGF-1-encoding DNA construct capable of expressing at least one human IGF-1 isoform, and a first pharmaceutically acceptable excipient; and

a second pharmaceutical composition comprising an HGF-encoding DNA construct capable of expressing at least one human HGF isoform, and a second pharmaceutically acceptable excipient.

64 - 121 . (canceled)