Method of enriching a target nucleic acid at a TNFRSF1B gene locus in a sample from a subject with inflammatory bowel disease
The aspects disclosed herein describe methods of identifying a subject that is non-responsive to anti-TNF therapy. The aspects disclosed herein further provide for a method of selecting a therapy for a subject with Inflammatory Bowel Disease (IBD), and treating the subject with the therapy.
1. A method of enriching a target nucleic acid in a sample, the method comprising:
(a) providing a biological sample from a subject with an inflammatory bowel disease, the biological sample comprising a target nucleic acid molecule at a TNFRSF1B gene locus comprising a cytosine at rs5745994 provided at nucleobase position 76 with reference to SEQ ID NO: 1;
(b) bringing a fluid reaction formulation comprising a synthetic oligonucleotide molecule in contact with the biological sample;
(c) hybridizing the synthetic oligonucleotide molecule and the target nucleic acid molecule at the TNFRSF1B gene locus comprising the cytosine at rs5745994;
(d) amplifying the target nucleic acid molecule at the TNFRSF1B gene locus comprising the cytosine at rs5745994, thereby enriching the target nucleic acid in the fluid reaction formulation; and
(e) detecting the enriched target nucleic acid molecule at the TNFRSF1B gene locus comprising the cytosine at rs5745994.
2. The method of claim 1 , wherein the inflammatory bowel disease comprises Crohn's disease (CD) or ulcerative colitis (UC).
3. The method of claim 1 , wherein detecting in (e) is indicative of the subject having a high risk of a non-response to an anti-Tumor Necrosis Factor alpha (TNFα) therapy.
4. The method of claim 1 , wherein the synthetic oligonucleotide comprises a detectable label, and wherein detecting in (e) is performed by detecting the label released from the synthetic oligonucleotide before amplification.
5. The method of claim 1 , wherein the biological sample further comprises a level of an antineutrophil cytoplasmic antibody (ANCA).
6. The method of claim 5 , further comprising detecting the level of ANCA by:
(a) bringing one or more antibodies in contact with a component of the biological sample comprising one or more antineutrophil cytoplasmic antibodies; and
(b) measuring a level of binding between the one or more antibodies and the one or more antineutrophil cytoplasmic antibodies.
7. The method of claim 6 , wherein the biological sample is blood, and the component of the biological sample is blood serum.
8. The method of claim 6 , wherein the level of binding between the one or more antibodies and the one or more antineutrophil cytoplasmic antibodies measured in (b) is high relative to a cutoff of greater than 2 standard deviations (SD) above mean control titers, when measured using a ELISA assay.
9. The method of claim 8 , wherein the level of binding between the one or more antibodies and the one or more antineutrophil cytoplasmic antibodies is indicative of the subject having a high risk of a non-response to an anti-TNFα therapy, relative to an individual with a level of binding between the one or more antibodies and the one or more antineutrophil cytoplasmic antibodies that is lower than the cutoff of 2 SD above mean control titers, when measured using the ELISA assay.
10. The method of claim 1 , further comprising:
(a) measuring a level of Tumor necrosis factor receptor 2 (TNFR2) in the biological sample; and
(b) detecting a high level of the TNFR2, relative to a level of TNFR2 in an individual that does not express the cytosine at rs5745994, provided at nucleobase position 76 with reference to SEQ ID NO: 1.
11. The method of claim 10 , wherein the level of TNFR2 that is high is indicative of the subject having a high risk of a non-response to an anti-TNFα therapy, relative to an individual that does not express the cytosine at rs5745994, provided at nucleobase position 76 with reference to SEQ ID NO: 1.
12. The method of claim 11 , further comprising treating the inflammatory bowel disease in the subject by administering to the subject a non-TNF therapy.
13. The method of claim 12 , wherein the non-TNF therapy comprises vedolizumab, ustekinumab, natalizumab, cyclosporine or Thalomide, or a combination thereof.
14. The method of claim 9 , further comprising treating the inflammatory bowel disease in the subject by administering to the subject a non-TNF therapy.
15. The method of claim 14 , wherein the non-TNF therapy comprises vedolizumab, ustekinumab, natalizumab, cyclosporine or Thalomide, or a combination thereof.