	US 12,220,416 B2
	Methylene-tetrahydrofolate reductase isoform-based biomarker and method of use thereof
Tibor A. Rauch, Oak Park, IL (US); Attila Balog, Szeged (HU); and Timea Ocsko, Chicago, IL (US)
Assigned to RUSH UNIVERSITY MEDICAL CENTER, Chicago, IL (US)
Filed by RUSH UNIVERSITY MEDICAL CENTER, Chicago, IL (US)
Filed on Jul. 19, 2018, as Appl. No. 16/040,204.
Claims priority of provisional application 62/534,972, filed on Jul. 20, 2017.
Prior Publication US 2019/0022099 A1, Jan. 24, 2019
Int. Cl. A61P 37/06 (2006.01); A61K 31/52 (2006.01); C12N 15/113 (2010.01); C12Q 1/6883 (2018.01); G01N 33/50 (2006.01)

CPC A61K 31/52 (2013.01) [A61P 37/06 (2018.01); C12N 15/1137 (2013.01); C12Q 1/6883 (2013.01); C12Q 2600/106 (2013.01); C12Q 2600/156 (2013.01); C12Q 2600/158 (2013.01); C12Q 2600/172 (2013.01); G01N 33/5091 (2013.01); G01N 2800/50 (2013.01); G01N 2800/52 (2013.01)]

8 Claims

1. A method for determining sensitivity in a subject to a treatment, the method comprising:

determining expression levels of a full-length MTHFR encoding mRNA and a truncated MTHFR encoding mRNA in a cell from the subject comprising:

(a) performing isoform-specific qRT-PCR of the full-length MTHFR encoding mRNA using a first primer consisting of SEQ ID NO 5 and a second primer consisting of SEQ ID NO 6,

(b) performing isoform-specific qRT-PCR of the truncated MTHFR encoding mRNA using a third primer consisting of SEQ ID NO 7 and a fourth primer consisting of SEQ ID NO 8,

c) calculating a Li/Si score using the Ct values generated in steps (a) and (b); and

determining the patient is not a candidate for treatment with one or more of methotrexate, azathioprine, chloroquine, or naltrexone;

administering a treatment comprising one or more anti-inflammatory drugs excluding methotrexate, azathioprine, chloroquine, or naltrexone;

wherein the Li/Si score is between a threshold value of 0.65 and 2.0.