Cell division marker

• US 8,420,306 B2
• Filed: 03/22/2010
• Issued: 04/16/2013
• Est. Priority Date: 10/06/2005
• Status: Active Grant

• Current Assignee: University Of Massachusetts
• Original Assignee: University Of Massachusetts
• Inventors: Doxsey, Stephen J., Chen, Chun-Ting
• Primary Examiner(s): Gabel, Gail R
• Application Number:
  US12/728,627
• Publication Number:
  US 20110117573A1
• Time in Patent Office: 1,121 Days
• Field of Search: 435/1.1, 435/6, 435/7.1, 435/7.2, 435/7.21, 435/7.23, 435/7.24, 435 792- 795, 435/40.5, 435/40.51, 435/40.52, 435/354, 435/358, 435/363, 435/367, 435/371, 435/372, 435/373, 435/374, 435/376, 435/976, 435/2, 435/347, 435/366, 436/512, 436/517, 436/519, 436/63, 436/64, 436/164, 436/172, 436/813
• US Class Current: 435/2
• 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 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;

detecting a cell having one or more midbody scars by a method comprising contacting the population of mammalian cells with an antibody or an antigen-binding portion thereof that binds specifically to a polypeptide component of a midbody scar selected from the group consisting of;

mitotic kinesin-like protein 1 (MKLP1), Epsilon tubulin (ε

-tubulin), Centrosomal Protein 55KDa (Cep55), and Aurora B kinase, and detecting binding of the antibody or antigen-binding portion thereof to the polypeptide component, wherein a ring structure not at a midbody or centrosome is the midbody scar; and

identifying the cell having one or more midbody scars as the dividing cell.

2. The method of claim 1, wherein the population of cells comprises dividing cells and non-dividing cells.

3. The method of claim 1, wherein the dividing cell is a cancerous cell.

4. The method of claim 1, wherein the dividing cell is a stem or progenitor cell.

5. The method of claim 1, further comprising identifying cells having a pre-selected number of midbody scars.

6. The method of claim 5, wherein the pre-selected number is 0 to 2 midbody scars per cell.

7. The method of claim 5, wherein the pre-selected number is 4 or more midbody scars per cell.

8. The method of claim 5, further comprising selecting cells having the pre-selected number of midbody scars.

9. The method of claim 8, further comprising ablating the selected cells.

10. The method of claim 8, further comprising isolating the selected cells.

11. The method of claim 1, wherein the detecting step comprises optically detecting the presence of a cellular structure having morphological characteristics of the midbody scar.

12. The method of claim 1, wherein optically detecting the presence of the cellular structure comprises using phase contrast microscopy to analyze the cell.

13. The method of claim 12, wherein the analysis is automated.

14. The method of claim 1, wherein the antibody or antigen binding portion thereof is a detectable antibody or antigen-binding portion thereof.

15. The method of claim 14 , wherein the detection is automated.

16. The method of claim 14 , wherein the detectible antibody comprises an enzyme having a detectable enzymatic activity.

17. The method of claim 14, wherein the detectible antibody or antigen binding portion thereof comprises a fluorescent label.

18. The method of claim 17 , wherein the detecting step comprises detecting the presence of fluorescence from the detectible antibody or antigen binding portion thereof.

19. The method of claim 18, wherein detecting the presence of fluorescence comprises using fluorescence microscopy.

20. The method of claim 1, wherein the population of cells is from a subject suspected of having a proliferative disorder.

21. The method of claim 20, further comprising:

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

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; and

identifying a subject having an elevated percentage of dividing cells in the sample as compared to the reference percentage as having a proliferative disorder.

22. The method of claim 20, 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.

23. The method of claim 1, further comprising isolating the dividing cells.

24. The method of claim 23, further comprising expanding the isolated dividing cells in culture.

25. The method of claim 1, wherein the ring structure is about 2μ

m in diameter.

26. The method of claim 1, wherein the ring structure is localized adjacent to the plasma membrane.

27. The method of claim 1, wherein the ring structure is determined not to comprise γ

-tubulin.

28. An isolated midbody scar from a mammalian cell, wherein the midbody scar is a ring structure not at a midbody or centrosome that comprises mitotic kinesin-like protein 1 (MKLP1), Epsilon tubulin (ε

-tubulin), Centrosomal Protein 55kDa (Cep55), and Aurora B kinase, and the midbody scar is isolated by a method that comprises contacting a mammalian cell with an antibod or an antigen-binding portion thereof that binds s ecificall to one of;

mitotic kinesin-like protein 1 (MKLP1), Epsilon tubulin (ε

-tubulin), Centrosomal Protein 55KDa (Cep55), and Aurora B kinase.