IP Library › Patent Application 10552485

Patent Application

App. No. 10/552,485

Lrp4/Corin Dopaminergic Neuron Progenitor Cell Markers

Loading inventors, assignments & file history…

Monitor This Case

Get email alerts when status or documents change.

Order Certified Copies

Most orders are placed with the USPTO same day — all within 24 business hours.

Order via The Patent Place →

Pre-filled with this patent's details

Quick Facts

Patent No.: US None
App. No.: 10/552,485

External Links

USPTO Patent Center↗ Full Patent PDF↗

Abstract

The present invention relates to polynucleotide probes and antibodies for detecting Lrp4/Corin dopaminergic neuron progenitor cell markers, which enable the efficient separation of dopaminergic neuron progenitor cells; and methods for selecting the progenitor cells by the use thereof.

Claims (125)

1 . A dopaminergic neuron proliferative progenitor cell marker polynucleotide probe comprising a sequence selected from the following nucleotide sequences (1) to (5):

(1) a nucleotide sequence complementary to a nucleotide sequence of SEQ ID NO: 1 or 2;

(2) a nucleotide sequence complementary to a nucleotide sequence encoding an amino acid sequence of SEQ ID NO: 3 or 4;

(3) a nucleotide sequence complementary to a nucleotide sequence encoding a sequence lacking a transmembrane domain in an amino acid sequence of SEQ ID NO: 3 or 4;

(4) a nucleotide sequence that hybridizes under stringent conditions with a polynucleotide consisting of a nucleotide sequence of SEQ ID NO: 1 or 2; and,

(5) a nucleotide sequence comprising at least 15 contiguous nucleotides selected from sequences of (1) to (4).

2 . A method for selecting a dopaminergic neuron proliferative progenitor cell, wherein the method comprises the step of contacting the polynucleotide probe of claim 1 with a cell sample thought to comprise a dopaminergic neuron proliferative progenitor cell.

3 . A method for selecting a dopaminergic neuron lineage cell, wherein the method comprises the steps of:

(1) selecting a dopaminergic neuron proliferative progenitor cell using the method of claim 2 for selecting the dopaminergic neuron proliferative progenitor cell;

(2) culturing the proliferative progenitor cell selected in step (1); and

(3) screening the cells cultured in step (2) by using a marker for a postmitotic dopaminergic neuron.

4 . A dopaminergic neuron proliferative progenitor cell, which is selected by the method of claim 2 .

5 . A method for isolating a dopaminergic neuron proliferative progenitor cell-specific gene and a gene specific for each maturation stage from the proliferative progenitor cell to a dopaminergic neuron, wherein the method comprises the step of detecting and isolating a gene specifically expressed in the proliferative progenitor cell of claim 4 or a cell which is differentiated, induced, or proliferated from the proliferative progenitor cell.

6 . A method of screening for a compound which regulates proliferation and/or differentiation of a dopaminergic neuron lineage cell using maturation as an index, wherein the method comprises the steps of: contacting a test substance with the proliferative progenitor cell of claim 4 or a cell which is differentiated, induced, or proliferated from the proliferative progenitor cell; and detecting a change of the proliferative progenitor cell or the progenitor cell caused by the contact.

7 . An antibody against a polypeptide selected from the following (1) to (6):

(1) a polypeptide encoded by a polynucleotide consisting of a nucleotide sequence of SEQ ID NO: 1 or 2;

(2) a polypeptide comprising an amino acid sequence of SEQ ID NO: 3 or 4;

(3) a polypeptide comprising an amino acid sequence lacking a transmembrane domain in an amino acid sequence of SEQ ID NO: 3 or 4;

(4) a polypeptide comprising an amino acid sequence with a deletion, insertion, substitution, or addition of one or more amino acids in an amino acid sequence of SEQ ID NO: 3 or 4;

(5) a polypeptide encoded by a polynucleotide that hybridizes under stringent conditions with a polynucleotide consisting of a sequence complementary to a nucleotide sequence of SEQ ID NO: 1 or 2; and,

(6) a polypeptide that is a fragment of a polypeptide of (1) to (5) comprising at least eight amino acid residues.

8 . The antibody of claim 7 , which is produced by the hybridoma FERM BP-10315 or FERM BP-10316.

9 . A dopaminergic neuron progenitor cell marker antibody, which comprises the antibody of claim 7 .

10 . A method for selecting a dopaminergic neuron progenitor cell, wherein the method comprises the step of contacting the antibody of claim 7 with a cell sample thought to comprise a dopaminergic neuron progenitor cell.

11 . A method for selecting a dopaminergic neuron lineage cell, wherein the method comprises the steps of:

(1) selecting a dopaminergic neuron proliferative progenitor cell using the method of claim 10 ;

(2) culturing the progenitor cell selected in step (1); and

(3) screening the progenitor cells cultured in step (2) by using a marker for a postmitotic dopaminergic neuron.

12 . A dopaminergic neuron progenitor cell, which is selected by the method of claim 10 .

13 . A method for isolating a dopaminergic neuron progenitor cell-specific gene and a gene specific for each maturation stage from the progenitor cell to a dopaminergic neuron, wherein the method comprises the step of detecting and isolating a gene specifically expressed in the progenitor cell of claim 12 or a cell which is differentiated, induced, or proliferated from the progenitor cell.

14 . A method of screening for a compound which regulates proliferation and/or differentiation of a dopaminergic neuron lineage cell using maturation as an index, wherein the method comprises the steps of: contacting a test substance with the progenitor cell of claim 12 or a cell which is differentiated, induced, or proliferated from the progenitor cell; and detecting a differentiated or proliferated progenitor cell caused by the contact.

15 . A kit for treating Parkinson's disease, which comprises the dopaminergic neuron proliferative progenitor cell of claim 4 or the dopaminergic neuron progenitor cell of claim 12 .

16 . A method for treating Parkinson's disease, wherein the method comprises the step of transplanting the dopaminergic neuron proliferative progenitor cell of claim 4 or the dopaminergic neuron progenitor cell of claim 12 into the brain of a patient.

17 . A use of the dopaminergic neuron proliferative progenitor cell of claim 4 or the dopaminergic neuron progenitor cell of claim 12 , for producing a kit for treating Parkinson's disease.

18 . A method for detecting or selecting a dopaminergic neuron proliferative progenitor cell, which comprises the step of contacting a cell sample comprising the dopaminergic neuron proliferative progenitor cell with a second polynucleotide which hybridizes under stringent conditions with a first polynucleotide consisting of any one of:

(1) the nucleotide sequence of SEQ ID NO: 1 or 2;

(2) a nucleotide sequence consisting of a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 3 or 4;

(3) a nucleotide sequence consisting of a polynucleotide encoding a polypeptide consisting of an amino acid sequence which lacks a transmembrane region in the amino acid sequence of SEQ ID NO: 3 or 4; and

(4) a nucleotide sequence consisting of a polynucleotide which hybridizes with a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1 or 2 under stringent conditions.

19 . The method of claim 18 , wherein the second polynucleotide comprises at least 15 nucleotides.

20 . A dopaminergic neuron proliferative progenitor cell population, which is selected by the method of claim 18 .

21 . A reagent for discriminating a dopaminergic neuron proliferative progenitor cell, which comprises a second polynucleotide as an active ingredient which hybridizes under stringent conditions with a first polynucleotide consisting of any one of:

(1) the nucleotide sequence of SEQ ID NO: 1 or 2;

(2) a nucleotide sequence consisting of a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 3 or 4;

(3) a nucleotide sequence consisting of a polynucleotide encoding a polypeptide comprising an amino acid sequence which lacks a transmembrane region in the amino acid sequence of SEQ ID NO: 3 or 4; and

(4) a nucleotide sequence consisting of a polynucleotide which hybridizes with a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1 or 2 under stringent conditions.

22 . The method of claim 21 , wherein the second polynucleotide comprises at least 15 nucleotides.

23 . A method for producing a postmitotic dopaminergic neuron precursor cell, wherein the method comprises the steps of:

(1) selecting a dopaminergic neuron proliferative progenitor cell by the method of claim 18 ;

(2) culturing the cell selected in step (1); and

(3) selecting the postmitotic dopaminergic neuron precursor cell from the cells cultured in step (2).

24 . A method for producing a dopaminergic neuron, wherein the method comprises the steps of:

(1) selecting a dopaminergic neuron proliferative progenitor cell by the method of claim 18 ; and

(2) culturing the cell selected in step (1).

25 . The method of claim 24 , further comprising the step of:

(3) selecting a dopaminergic neuron from the cells cultured in step (2).

26 . A method for detecting or selecting a dopaminergic neuron progenitor cell, which comprises the step of contacting a cell sample comprising the dopaminergic neuron progenitor cell with an antibody which is bound to a polypeptide consisting of the amino acid sequence of any one of:

(1) the amino acid sequence of SEQ ID NO: 3 or 4;

(2) an amino acid sequence which lacks a transmembrane region in the amino acid sequence of SEQ ID NO: 3 or 4;

(3) an amino acid sequence mutated by one or more amino acid deletions, substitutions, or additions, or any combination thereof, in the amino acid sequence of SEQ ID NO:3 or 4; and

(4) an amino acid sequence consisting of a polypeptide encoded by a polynucleotide which hybridizes under stringent conditions with a polynucleotide consisting of a nucleotide sequence complementary to a nucleotide sequence of SEQ ID NO: 1 or 2,

or a partial sequence thereof.

27 . The method of claim 26 , wherein the polypeptide comprising the partial sequence comprises at least six consecutive amino acid residues.

28 . A dopaminergic neuron progenitor cell population, which is selected by the method of claim 26 .

29 . The cell population of claim 28 , which comprises 40% or more dopaminergic neuron progenitor cells in the entire cells.

30 . A reagent for discriminating a dopaminergic neuron progenitor cell, which comprises an antibody as an active ingredient which is bound to a polypeptide consisting of the amino acid sequence of any one of:

(1) the amino acid sequence of SEQ ID NO: 3 or 4;

(2) an amino acid sequence which lacks a transmembrane region in the amino acid sequence of SEQ ID NO: 3 or 4;

(3) an amino acid sequence mutated by one or more amino acid deletions, substitutions, or additions, or any combination thereof, in the amino acid sequence of SEQ ID NO:3 or 4; and

31 . The reagent of claim 30 , wherein the polypeptide consisting of the partial sequence comprises at least six consecutive amino acid residues.

32 . The reagent of claim 30 , wherein the antibody is produced by the hybridoma FERM BP-10315 or FERM BP-10316.

33 . An antibody produced by the hybridoma FERM BP-10315 or FERM BP-10316.

34 . A method for producing a dopaminergic neuron proliferative progenitor cell, wherein the method comprises the steps of:

(1) selecting a dopaminergic neuron progenitor cell by the method of claim 26 ; and

(2) removing a postmitotic dopaminergic neuron precursor cell to select the dopaminergic neuron proliferative progenitor cell.

35 . A method for producing a postmitotic dopaminergic neuron precursor cell, wherein the method comprises the steps of:

(1) selecting a dopaminergic neuron progenitor cell by the method of claim 26 ; and

(2) culturing the cell selected in step (1).

36 . The method of claim 35 , further comprising the step of:

(3) selecting a postmitotic dopaminergic neuron precursor cell from the cells cultured in step (2).

37 . A method for producing a dopaminergic neuron, wherein the method comprises the steps of:

(1) selecting a dopaminergic neuron progenitor cell by the method of claim 26 ; and

(2) culturing the cell selected in step (1).

38 . The method of claim 37 , further comprising the step of:

(3) selecting a dopaminergic neuron from the cells cultured in step (2).

39 . A kit for treating a neurodegenerative disease, which comprises at least one cell selected from the group consisting of:

(1) the dopaminergic neuron proliferative progenitor cell population of claim 20 ;

(2) a postmitotic dopaminergic neuron precursor cell produced by the method of claim 23 ;

(3) a dopaminergic neuron produced by the method of claim 24 ;

(4) a dopaminergic neuron produced by the method of claim 25 ;

(5) the dopaminergic neuron progenitor cell population of claim 28 ;

(6) the dopaminergic neuron progenitor cell population of claim 29 ;

(7) a dopaminergic neuron proliferative progenitor cell produced by the method of claim 34 ;

(8) a postmitotic dopaminergic neuron precursor cell produced by the method of claim 35 ;

(9) a postmitotic dopaminergic neuron precursor cell produced by the method of claim 36 ;

(10) a dopaminergic neuron produced by the method of claim 37 ; and

(11) a dopaminergic neuron produced by the method of claim 38 .

40 . The kit of claim 39 , wherein the neurodegenerative disease is Parkinson's disease.

41 . A method for treating a neurodegenerative disease, which comprises the step of transplanting into the brain of a patient at least one cell selected from the group consisting of: