Diagnostic methods and kits for monitoring response to chemotherapy in ovarian cancer
AssigneesRNA Disgnostics Inc., Laurentian University of Sudbury
InventorsAmadeo Mark Parissenti, Baoqing Guo, Kenneth Pritzker, Laura Pritzker
Life of this patent
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Abstract
Provided are methods of determining a response to a chemotherapeutic agent in a subject with ovarian cancer, comprising: determining a RNA integrity value of a sample comprising ovarian cancer cell RNA from the subject after the subject has received one or more doses of the chemotherapeutic agent; wherein a low RNA integrity value and/or RNA degradation of the cancer cell RNA is indicative that the cancer is responding to the chemotherapeutic agent and/or a high RNA integrity value and/or stable RNA integrity of the ovarian cancer cell RNA is indicative that the cancer is resistant to the chemotherapeutic agent.
Claims12
An in vitro method of identifying synergistic combinations of chemotherapeutic agents, comprising:
- a. harvesting human ovarian cancer cells, placing the harvested cells in different test wells, and incubating the harvested cells in the different test wells with each chemotherapeutic agent and with a combination of the chemotherapeutic agents for at least 1 hour;
- b. collecting floating and adherent ovarian cancer cells for each of the test wells;
- c. isolating RNA from each of the collected ovarian cancer cells to provide a RNA sample for each test well, said isolating comprising lysing the collected ovarian cancer cells in a RNA stabilizing composition, preparing a precipitation mixture of the lysed ovarian cancer cells, adding the precipitation mixture to a RNA isolating column and eluting the RNA from the column with a RNAse free liquid;
- d. determining a RNA integrity value of each RNA sample, said determining comprising separating the RNA sample using capillary electrophoresis apparatus;
- producing an electropherogram of the separated RNA sample and quantitating the RNA integrity of the separated RNA sample by assessing the electropherogram, wherein the RNA integrity value is a RNA integrity number (RIN);
- e. identifying the chemotherapeutic combination as synergistic when the reduction in RNA integrity is greater than any additive reduction in RNA integrity of the cancer cells treated with each agent of the combination alone or otherwise identifying the combination as non-synergistic;
- and
- f. testing the synergistic combinations in one or more assays selected from a clonogenic assay.
Dependencies are inferred from the claim text.