	US 12,253,524 B2
	Methods and compositions for determining the functional activity of DNA double strand break repair pathway molecules for assessing germline risk of cancer
Harry Ostrer, New York, NY (US); Johnny C. Loke, Nanuet, NY (US); and Alexander Pearlman, New York, NY (US)
Assigned to Albert Einstein College of Medicine, Bronx, NY (US)
Filed by ALBERT EINSTEIN COLLEGE OF MEDICINE, Bronx, NY (US)
Filed on Apr. 6, 2023, as Appl. No. 18/131,559.
Application 18/131,559 is a continuation of application No. 16/900,194, filed on Jun. 12, 2020, granted, now 11,650,206.
Application 16/900,194 is a continuation of application No. 15/504,726, granted, now 10,718,774, issued on Jul. 21, 2020, previously published as PCT/US2015/045856, filed on Aug. 19, 2015.
Claims priority of provisional application 62/039,691, filed on Aug. 20, 2014.
Prior Publication US 2023/0258646 A1, Aug. 17, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. G01N 33/574 (2006.01); C12Q 1/6886 (2018.01)

CPC G01N 33/57496 (2013.01) [C12Q 1/6886 (2013.01); G01N 33/57415 (2013.01); G01N 33/57449 (2013.01); G01N 33/57484 (2013.01); C12Q 2600/156 (2013.01); G01N 2333/4703 (2013.01); G01N 2333/4748 (2013.01); G01N 2333/9108 (2013.01); G01N 2440/14 (2013.01); G01N 2800/50 (2013.01)]

16 Claims

1. A method comprising:

obtaining circulating white blood cells from a subject;

treating the white blood cells with a DNA damaging agent;

performing a flow cytometry based functional variant analysis (FVA);

measuring in the treated cell at least one functional activity of a DNA double strand break (DSB) repair pathway gene comprising ATM, BRCA1, BRCA2, PALB2, FANCD2, FANCC, FANCF, NBN, BARD1, p53, RAD50/51, NBS1, Abraxas, CtIP, and DNA Ligase genes;

comparing the at least one measured functional activity with at least one control value obtained from control white blood cells treated with the DNA damaging agent and having a wild type DNA double strand break (DSB) repair pathway; and

categorizing DSB repair pathway gene in the subject as functional, having loss of function or having a gain of function, based on the comparing step, wherein the DSB repair pathway gene comprises ATM, BRCA1, BRCA2, PALB2, FANCD2, FANCC, FANCF, NBN, BARD1, p53, RAD50/51, NBS1, Abraxas, CtIP, and DNA Ligase genes.