US 12,331,345 B2
Methods and compositions for diagnosing and treating, germline mismatch repair deficiencies, lynch syndrome and assessing germline risks of cancer
Harry Ostrer, New York, NY (US); Johnny C. Loke, Valley Cottage, NY (US); and Ishraq Alim, New York, NY (US)
Filed by MORGAN AND MENDEL GENOMICS, INC., New York, NY (US); and ALBERT EINSTEIN COLLEGE OF MEDICINE, Bronx, NY (US)
Filed on Feb. 17, 2022, as Appl. No. 17/674,400.
Claims priority of provisional application 63/233,064, filed on Aug. 13, 2021.
Claims priority of provisional application 63/150,347, filed on Feb. 17, 2021.
Prior Publication US 2022/0260553 A1, Aug. 18, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. G01N 31/00 (2006.01); C12Q 1/6804 (2018.01); C12Q 1/6827 (2018.01); G01N 15/14 (2006.01); G01N 33/50 (2006.01); G01N 33/53 (2006.01); G01N 33/574 (2006.01); G01N 15/10 (2006.01)
CPC C12Q 1/6827 (2013.01) [C12Q 1/6804 (2013.01); G01N 15/14 (2013.01); G01N 33/5023 (2013.01); G01N 33/5035 (2013.01); G01N 33/5041 (2013.01); G01N 33/57419 (2013.01); G01N 33/57484 (2013.01); G01N 2015/1006 (2013.01)] 6 Claims
 
1. A method for screening a population of cells from a human subject for functionality of the mismatch repair (MMR) pathway, the method comprising:
(a) contacting the population of cells with a DNA mismatch-inducing agent;
(b) measuring via flow-cytometry in the population of cells at least one functional activity of the mismatch repair (MMR) pathway, wherein the at least one functional activity comprises nuclear localization of mutL homologue 1 (MLH1) and mutS homologue 2 (MSH2);
(c) comparing the nuclear localization of MLH1 and MSH2 in the population of cells contacted with the DNA mismatch-inducing agent with the nuclear localization of MLH1 and MSH2 from a control population of cells treated with the DNA mismatch-inducing agent and having a normal functioning MMR pathway; and
(d) categorizing the MMR pathway as having loss of function or as having a partial-loss of function, based on the comparing step (c);
wherein having loss of function or partial-loss of function of the MMR pathway indicates the subject as having Lynch Syndrome and one or more preventative interventions selected from colonoscopy, chemotherapy, chemoprevention, radiotherapy, immunotherapy, gene therapy, or surgery is administered to the subject having Lynch Syndrome.