Methods for reducing and/or preventing excessive cellular apoptosis

a cell death and cell technology, applied in the field of reducing and/or preventing excessive cellular apoptosis, can solve the problems of apoptosis in some cells, apoptosis death in some cells, apoptosis death, etc., to prevent or reduce the number of cardiomyocytes, prevent or reduce the number of apoptosis-induced cell death, and reduce the effect of oxygen delivery

Inactive Publication Date: 2014-02-06
STEMNION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0005]Many pathological conditions feature excessive cellular apoptosis (i.e., neurodegenerative diseases, AIDS, ischemia, radiation exposure, to name a few) and thus may benefit from therapeutically preventing or reducing apoptosis-induced cell death. For example, excessive cellular apoptosis is thought to play an important role in myocardial ischemia caused by an insufficient blood supply, leading to a decrease in oxygen delivery to, and subsequent death of, the cardiomyocytes. The methods of the invention may be useful to prevent or reduce the number of cardiomyocytes undergoing apoptosis-induced death.

Problems solved by technology

Irradiation or drugs used for cancer chemotherapy results in DNA damage in some cells, which can lead to apoptotic death through a p53-dependent pathway.
Some hormones, such as corticosteroids, may lead to apoptotic death in some cells (e.g., thymocytes) although other cells are unaffected or even stimulated.
For example, excessive cellular apoptosis is thought to play an important role in myocardial ischemia caused by an insufficient blood supply, leading to a decrease in oxygen delivery to, and subsequent death of, the cardiomyocytes.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

Evaluation of the Effect of ACCS on Apoptosis in Jurkat Cells

[0079]Apoptosis is programmed cell death and is a necessary event for proper healing and tissue turnover involving a complex network of biochemical pathways. Dis-regulation of apoptosis is a cause of delayed wound healing as well as many chronic diseases.

[0080]Depending on the extent of damage, cells will either apoptose or survive. Applicants have discovered that ACCS contains anti-apoptotic factors that may help some cells survive various insults that would typically lead to apoptosis. From a therapeutic perspective, these anti-apoptotic factors may benefit partially damaged cells such as those found in a wound by helping them survive and repopulate the wound bed quicker.

[0081]Initial experiments were performed using a FITC Annexin V staining kit (BD Biosciences) and analyzed by Flow Cytometry.

[0082]Brief description of the assay: in apoptotic cells, the membrane phospholipid phosphatidylserine (PS) is translocated from ...

example 2

Evaluation of the Effect of ACCS on Apoptosis in Human Foreskin Fibroblasts Cells

[0085]Additional experiments were performed using an apoptosis assay kit which tests for the activation of both caspase 3 and caspase 7, both of which are key biomarkers of apoptosis.

[0086]Brief description of the assay: The Caspase-Glo® 3 / 7 Assay (Promega) provides a luminogenic caspase-3 / 7 substrate, which contains the tetrapeptide sequence DEVD, in a reagent optimized for caspase activity, luciferase activity and cell lysis. Following caspase cleavage, a substrate for luciferase (aminoluciferin) is released, resulting in the luciferase reaction and the production of light. The higher the luminescence the more caspase activity in the sample.

[0087]In this particular experiment, Human Foreskin Fibroblasts (HFF's) (˜20,000 cells / well) were plated in 96-well plates and then treated for 5 hrs with and without Staurosporine, an apoptosis inducer, in control media, ACCS, irradiated ACCS, or normal growth med...

example 3

Evaluation of the Effects of ACCS on Radiations Protection in Jurkat Cells

[0089]Based on the above results, it was hypothesized that ACCS may protect cells from exposure to radiation, a known apoptosis inducer. Experiments were performed using Jurkat Cells to evaluate the potential of ACCS as a radioprotection agent. In one such experiment, in a total volume of 1 mL, 2×106 Jurkat cells were irradiated at 1000, 2000 and 4000 RADs and apoptosis was analyzed by flow cytometry with Annexin V and the caspase luminescence assays. Treatments included control medium at 0.75× and ACCS at 0.75×. All treatments were added immediately prior to irradiating the cell samples.

[0090]Results—The results indicate that ACCS protects cells from ionizing radiation as indicated by a reduction (6.6, 19.4 and 18.2% reduction, respectively, for each irradiation dose as compared to control) in the percentage of apoptotic cells and a reduction (48, 42 and 25%, respectively, in caspase activity as compared to m...

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Abstract

The invention is directed to novel methods for reducing the number of apoptotic cell deaths in a population of cells undergoing excessive cellular apoptosis. The invention is also directed to novel methods for preventing apoptotic cell death in a population of cells at risk for developing excessive cellular apoptosis. In particular, the invention is directed to novel methods for reducing or preventing excessive cellular apoptosis comprising exposing cells exhibiting or at risk for developing excessive cellular apoptosis to a novel cellular factor-containing composition called Amnion-derived Cellular Cytokine Solution (referred to herein as ACCS), which is obtained from the culturing of Amnion-derived Multipotent Progenitor (AMP) cells, or AMP cells.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims priority under 35 USC §119(e) of U.S. Provisional Application No. 61 / 679,150, filed Aug. 3, 2012 and U.S. Provisional Application No. 61 / 768,429, filed Feb. 23, 2012, the entireties of which are incorporated herein by reference.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT[0002]This invention was made in part with United States government support awarded by the following agency: Naval Medical Logistics Command, Contract # N62645-11-1-001. The United States may have certain rights to this invention.FIELD OF THE INVENTION[0003]The field of the invention is directed to novel methods for reducing the number of apoptotic cell deaths in a population of cells undergoing excessive cellular apoptosis. The field of the invention is also directed to novel methods for preventing apoptotic cell death in a population of cells at risk for developing excessive cellular apoptosis. In particular, the field of the i...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N5/073
CPCC12N5/0605A61K38/1793A61K35/12A61K38/1866A61K2300/00C12N5/0636C12N5/0647C12N5/0656C12N2501/135C12N2501/15C12N2501/165C12N2501/40C12N2501/48C12N2501/73C12N2502/025
Inventor WESSEL, HOWARD C.BANAS, RICHARD A.
Owner STEMNION
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