Cell division marker

• US 20070154957A1
• Filed: 10/06/2006
• Published: 07/05/2007
• Est. Priority Date: 10/06/2005
• Status: Active Grant

• Current Assignee: University Of Massachusetts
• Original Assignee: University Of Massachusetts
• Inventors: Chen, Chun-Ting, Doxsey, Stephen
• Granted Patent: US 7,682,799 B2
• Time in Patent Office: Days
• US Class Current: 435/7.200
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: G01N 2800/085 Liver diseases, e.g. portal hypertension, fibrosis, cirrhosis, bilirubin
• CPC Class Codes: G01N 2800/12 Pulmonary diseases
• CPC Class Codes: G01N 2800/20 Dermatological disorders
• CPC Class Codes: G01N 2800/323 Arteriosclerosis, Stenosis
• CPC Class Codes: G01N 2800/328 Vasculitis, i.e. inflammation of blood vessels
• CPC Class Codes: G01N 2800/342 Prostate diseases, e.g. BPH, prostatitis
• CPC Class Codes: G01N 2800/347 Renal failures; Glomerular diseases; Tubulointerstitial diseases, e.g. nephritic syndrome, glomerulonephritis; Renovascular diseases, e.g. renal artery occlusion, nephropathy
• CPC Class Codes: G01N 2800/36 Gynecology or obstetrics
• CPC Class Codes: G01N 2800/364 Endometriosis, i.e. non-malignant disorder in which functioning endometrial tissue is present outside the uterine cavity
• CPC Class Codes: G01N 33/564 for pre-existing immune complex or autoimmune disease , i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9
• CPC Class Codes: G01N 33/56966 Animal cells
• CPC Class Codes: G01N 33/57484 involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
• CPC Class Codes: G01N 33/57492 involving compounds localized on the membrane of tumor or cancer cells

Abstract

This application relates to a newly identified animal cell structure, the midbody scar. This structure is a remnant of the midbody that is retained by one daughter cell following cytokinesis and persists through multiple subsequent cell cycles. The midbody scar can be useful as a marker of dividing cells or of a cell'"'"'s replicative age.

Claims

1. A method of detecting the presence of a midbody scar in a mammalian cell, the method comprising:

contacting a mammalian cell suspected to have a midbody scar with an antibody or antigen-binding portion thereof that binds specifically to a polypeptide component of a midbody scar selected from the group consisting of; and

detecting binding of the antibody to its antigen, wherein binding of the antibody indicates the presence of a midbody scar in the mammalian cell.

2. The method of claim 1, wherein the polypeptide is selected from the group consisting of mitotic kinesin-like protein 1 (MKLP1);

Epsilon tubulin (ε

-tubulin);

Centrosomal Protein 55 kDa (Cep55); and

Aurora B kinase.

3. The method of claim 1, comprising:

contacting the mammalian cell with two or more antibodies or antigen-binding portions thereof that each bind specifically to a polypeptide component of a midbody scar, and

detecting binding of the two or more antibodies,

wherein colocalized binding of the two or more of antibodies indicates the presence of a midbody scar.

5. The method of claim 1, wherein the cell is in interphase.

6. The method of claim 1, wherein the mammalian cell is a fixed cell.

7. The method of claim 6, wherein the fixed cell is in a pathology specimen.

8. The method of claim 1, wherein the cell is in tissue.

9. The method of claim 1, wherein the antibody or antigen-binding portion thereof is bound or linked to a detectable moiety.

10. The method of claim 9, wherein detecting binding of the antibody to its antigen comprises detecting the detectable moiety.

4. The method of claim 4, wherein the polypeptide is selected from the group consisting of mitotic kinesin-like protein 1 (MKLP1);

Epsilon tubulin (ε

-tubulin);

Centrosomal Protein 55 kDa (Cep55); and

Aurora B kinase.

11. A method of identifying a dividing cell in a population of mammalian cells, the method comprising:

obtaining a population of mammalian cells comprising at least one cell suspected of being a dividing cell; and

detecting a cell having one or more midbody scars, wherein a cell having one or more midbody scars is identified as a dividing cell.

12. The method of claim 11, wherein the population of cells comprises dividing cells and non-dividing cells.

13. The method of claim 11, wherein the dividing cell is a cancerous cell.

14. The method of claim 11, wherein the dividing cell is a stem or progenitor cell.

15. The method of claim 11, further comprising identifying cells having a desired number of midbody scars.

16. The method of claim 15, wherein the desired number is 0 to 2 midbody scars per cell.

17. The method of claim 15, wherein the desired number is 4 or more midbody scars per cell.

18. The method of claim 15, further comprising selecting cells having the desired number of midbody scars.

19. The method of claim 18, further comprising ablating the selected cells.

20. The method of claim 18, further comprising isolating the selected cells.

21. The method of claim 11, wherein identifying a cell with one or more midbody scars comprises optically detecting the presence of a cellular structure having morphological characteristics of a midbody scar.

22. The method of claim 21, wherein optically detecting the presence of a cellular structure comprises using phase contrast microscopy to analyze the cell.

23. The method of claim 22, wherein the analysis is automated.

24. The method of claim 11, wherein identifying a cell with one or more midbody scars comprises contacting the population of cells with a detectible antibody or antigen-binding portion thereof that binds specifically to a polypeptide component of a midbody scar;

and

detecting binding of the detectible antibody to the polypeptide, wherein binding of the antibody indicates the presence of a midbody scar.

25. The method of claim 24, wherein the polypeptide component of a midbody scar is selected from the group consisting of mitotic kinesin-like protein 1 (MKLP1);

Epsilon tubulin (ε

-tubulin);

Centrosomal Protein 55 kDa (Cep55); and

Aurora B kinase.

26. The method of claim 24, wherein the detectible antibody or antigen binding portion thereof comprises a fluorescent label.

27. The method of claim 26, wherein identifying a cell with one or more midbody scars comprises detecting the presence of fluorescence from the detectible antibody or antigen binding portion thereof.

28. The method of claim 27, wherein detecting the presence of fluorescence comprises using fluorescence microscopy.

29. The method of claim 24, wherein the detection is automated.

30. The method of claim 24, wherein the detectible antibody comprises an enzyme, which when provided with an appropriate substrate produces a product that is detected.

31. The method of claim 11, wherein the population of cells is from a subject suspected of having a proliferative disorder.

32. The method of claim 31, further comprising:

determining the percentage of cells in the sample that have a selected number of midbody scars; and

comparing the percentage of cells in the sample to a reference percentage representing a sample from a subject that does not have a proliferative disorder;

wherein the presence of a significantly higher number of dividing cells as compared to the reference sample indicates that the subject has a proliferative disorder.

33. The method of claim 31, wherein the proliferative disorder is selected from the group consisting of atherosclerosis, rheumatoid arthritis, idiopathic pulmonary fibrosis, scleroderma, cirrhosis of the liver, lupus, vasculitis, endometriosis, uterine fibroids, prostatic hyperplasia, nephritis, skin disorders, and cancer.

34. The method of claim 11, further comprising isolating the dividing cells.

35. The method of claim 34, further comprising expanding the isolated dividing cells in culture under conditions sufficient to provide a population of dividing cells.

36. An isolated midbody scar from a mammalian cell, wherein the midbody scar comprises mitotic kinesin-like protein 1 (MKLP1);

Epsilon tubulin (ε

-tubulin);

Centrosomal Protein 55 kDa (Cep55); and

Aurora B kinase.