Compositions for detecting cell death and methods of use thereof

• US 8,999,661 B2
• Filed: 07/29/2009
• Issued: 04/07/2015
• Est. Priority Date: 07/30/2008
• Status: Active Grant

• Current Assignee: Dana-Farber Cancer Institute
• Original Assignee: Dana-Farber Cancer Institute
• Inventors: Kung, Andrew, Banerjee, Pallab
• Primary Examiner(s): RAMIREZ, DELIA M
• Application Number:
  US12/511,478
• Publication Number:
  US 20100034800A1
• Time in Patent Office: 2,078 Days
• Field of Search: 435/190, 435/69.7, 435/26, 530/350
• US Class Current: 435/26
• CPC Class Codes: C12Q 1/32 involving dehydrogenase
• CPC Class Codes: G01N 2510/00 Detection of programmed cell death, i.e. apoptosis
• CPC Class Codes: G01N 33/5011 for testing antineoplastic activity

Abstract

The invention provides compounds and methods of their use in the detection of apoptosis and necrosis both in vitro and in vivo. Also provided are compounds and methods of their use in selective delivery of agents to cells undergoing apoptosis or necrosis. The compounds and methods are based on conjugates formed with a dehydrogenase such as lactate dehydrogenase, alcohol dehydrogenase, aldehyde dehydrogenase, and malate dehydrogenase. The compounds and methods are useful in the diagnosis and treatment of conditions characterized by apoptosis, including cancer, cardiac disease, neurologic disease including stroke, and autoimmunity. The compounds and methods offer distinct advantages over corresponding compounds and methods based on Annexin V. Also provided are methods for screening for compounds that modulate, i.e., inhibit or promote, apoptosis.

Claims

1. A method for detecting cells undergoing early apoptosis cell death, the method comprising:

contacting a test population of cells with a conjugate of a dehydrogenase linked to a detectable label, under conditions suitable for uptake of the conjugate by the test population of cells, wherein the dehydrogenase is a lactate dehydrogenase (LDH), alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH) or malate dehydrogenase (MDH); and

detecting the label in the test population of cells,

wherein presence of the label in the test population of cells indicates cells undergoing early apoptosis.

2. The method of claim 1, wherein the lactate dehydrogenase (LDH) is LDH-A, LDH-B or LDH-C.

3. The method of claim 2, wherein the LDH is a human LDH.

4. The method of claim 1, wherein the dehydrogenase is a human dehydrogenase.

5. The method of claim 1, wherein the contacting and the detecting are carried out in vitro.

6. The method of claim 1, wherein the contacting and the detecting are carried out ex vivo.

7. The method of claim 1, wherein at least one of the contacting and the detecting is carried out in vivo.

8. The method of claim 7, wherein the contacting and the detecting are carried out in vivo.

9. The method of claim 1, wherein the test population of cells is obtained from a subject having or suspected of having a condition associated with apoptosis.

10. The method of claim 9, wherein the condition is selected from the group consisting of:

cancer;

chemotherapy-, radiation-, or hormone-induced apoptosis in solid and hematological tumors;

tumor resistance to therapy;

acute cardiac allograft rejection;

acute myocardial infarction;

anthracycline-induced cardiotoxicity;

arrhythmogenic right ventricle dysplasia;

skeletal muscle apoptosis;

congestive heart failure;

coronary artery disease;

atherosclerosis;

infectious endocarditis;

myocarditis;

myocardial dysfunction;

myocardial ischemia-reperfusion injury;

non-cardiac allograft rejection;

bacterial infection;

viral infection;

multiple organ dysfunction syndrome;

septic shock;

cerebral ischemia-reperfusion injury;

macular degeneration;

neurodegenerative disease;

central nervous system trauma;

autoimmune diabetes mellitus;

rheumatoid arthritis;

systemic lupus erythematosus;

inflammatory bowel disease;

multiple sclerosis;

autoimmune diseases;

annexinopathies;

osteoarthritis;

renal failure;

chronic renal atrophy and renal fibrosis;

glomerular injury; and

polycystic renal disease.

11. The method of claim 1, further comprising:

measuring the amount of label detected in the test population of cells;

comparing the amount of label detected in the test population of cells to a reference amount of label detected in a reference population of cells; and

determining if (a) there is an increased amount of apoptosis when the amount of label detected in the test population of cells is greater than the reference amount of detected label, or (b) there is a decreased amount of apoptosis when the amount of label detected in the test population of cells is less than the reference amount of detected label.