title: Combined genetic effects of EGLN1 and VWF modulate thrombotic outcome in hypoxia revealed by Ayurgenomics approach.
abstract: BACKGROUND
  Extreme constitution "Prakriti" types of Ayurveda exhibit systemic physiological attributes. Our earlier genetic study has revealed differences in EGLN1, key modulator of hypoxia axis between Prakriti types. This was associated with differences in high altitude adaptation and susceptibility to high altitude pulmonary edema (HAPE). In this study we investigate other molecular differences that contribute to systemic attributes of Prakriti that would be relevant in predictive marker discovery.
  
  
  METHODS
  Genotyping of 96 individuals of the earlier cohort was carried out in a panel of 2,800 common genic SNPs represented in Indian Genomic Variation Consortium (IGVC) panel from 24 diverse populations. Frequency distribution patterns of Prakriti differentiating variations (FDR correction P < 0.05) was studied in IGVC and 55 global populations (HGDP-CEPH) panels. Genotypic interactions between VWF, identified from the present analysis, and EGLN1 was analyzed using multinomial logistic regression in Prakriti and Indian populations from contrasting altitudes. Spearman's Rank correlation was used to study this genotypic interaction with respect to altitude in HGDP-CEPH panel. Validation of functional link between EGLN1 and VWF was carried out in a mouse model using chemical inhibition and siRNA studies.
  
  
  RESULTS
  Significant differences in allele frequencies were observed in seven genes (SPTA1, VWF, OLR1, UCP2, OR6K3, LEPR, and OR10Z1) after FDR correction (P < 0.05). A non synonymous variation (C/T, rs1063856) associated with thrombosis/bleeding susceptibility respectively, differed significantly between Kapha (C-allele) and Pitta (T-allele) constitution types. A combination of derived EGLN1 allele (HAPE associated) and ancestral VWF allele (thrombosis associated) was significantly high in Kapha group compared to Pitta (p < 10(-5)). The combination of risk-associated Kapha alleles was nearly absent in natives of high altitude. Inhibition of EGLN1 using (DHB) and an EGLN1 specific siRNA in a mouse model lead to a marked increase in vWF levels as well as pro-thrombotic phenotype viz. reduced bleeding time and enhanced platelet count and activation.
  
  
  CONCLUSIONS
  We demonstrate for the first time a genetic link between EGLN1 and VWF in a constitution specific manner which could modulate thrombosis/bleeding susceptibility and outcomes of hypoxia. Integration of Prakriti in population stratification may help assemble common variations in key physiological axes that confers differences in disease occurrence and patho-phenotypic outcomes.
authors.affiliation: Genomics and Molecular Medicine, CSIR-Institute of Genomics and Integrative Biology, Sukhdev Vihar, Mathura Road, New Delhi, India. shilpi.aggarwal@igib.in.
authors.forename: Shilpi
authors.initials: S
authors.lastname: Aggarwal
authors.affiliation: CSIR's Ayurgenomics Unit-TRISUTRA (Translational Research and Innovative Science ThRough Ayurgenomics), CSIR-Institute of Genomics and Integrative Biology, Sukhdev Vihar, Mathura Road, New Delhi, 110 020, India. atish.gheware@igib.in.|Academy of Scientific and Innovative Research (AcSIR), New Delhi, India. atish.gheware@igib.in.
authors.forename: Atish
authors.initials: A
authors.lastname: Gheware
authors.affiliation: Genomics and Molecular Medicine, CSIR-Institute of Genomics and Integrative Biology, Sukhdev Vihar, Mathura Road, New Delhi, India. a.agrawal@igib.res.in.|Academy of Scientific and Innovative Research (AcSIR), New Delhi, India. a.agrawal@igib.res.in.
authors.forename: Anurag
authors.initials: A
authors.lastname: Agrawal
authors.affiliation: Indian Statistical Institute, Kolkata, India. saurabh@isical.ac.in.
authors.forename: Saurabh
authors.initials: S
authors.lastname: Ghosh
authors.affiliation: CSIR's Ayurgenomics Unit-TRISUTRA (Translational Research and Innovative Science ThRough Ayurgenomics), CSIR-Institute of Genomics and Integrative Biology, Sukhdev Vihar, Mathura Road, New Delhi, 110 020, India. bhavana.p@igib.res.in.|Academy of Scientific and Innovative Research (AcSIR), New Delhi, India. bhavana.p@igib.res.in.
authors.forename: Bhavana
authors.initials: B
authors.lastname: Prasher
authors.affiliation: Genomics and Molecular Medicine, CSIR-Institute of Genomics and Integrative Biology, Sukhdev Vihar, Mathura Road, New Delhi, India. mitali@igib.res.in.|CSIR's Ayurgenomics Unit-TRISUTRA (Translational Research and Innovative Science ThRough Ayurgenomics), CSIR-Institute of Genomics and Integrative Biology, Sukhdev Vihar, Mathura Road, New Delhi, 110 020, India. mitali@igib.res.in.|Academy of Scientific and Innovative Research (AcSIR), New Delhi, India. mitali@igib.res.in.
authors.forename: Mitali
authors.initials: M
authors.lastname: Mukerji
mesh_terms: D000222:Adaptation, Physiological / Q000235:genetics; D000483:Alleles; D000531:Altitude; D000818:Animals; D004195:Disease Models, Animal; D005787:Gene Frequency / Q000235:genetics; D055785:Gene Knockdown Techniques; D020022:Genetic Predisposition to Disease; D023281:Genomics; D006720:Homozygote; D006801:Humans; D000860:Hypoxia / Q000097:blood / Q000150:complications / Q000235:genetics*; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases / Q000235:genetics* / Q000378:metabolism; D007194:India; D008297:Male; D008515:Medicine, Ayurvedic; D008807:Mice, Inbred BALB C; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D015203:Reproducibility of Results; D013927:Thrombosis / Q000097:blood / Q000150:complications / Q000235:genetics*; D014841:von Willebrand Factor / Q000235:genetics*
chemical_list: D014841:von Willebrand Factor; C490190:EGLN1 protein, human; C490189:Egln1 protein, mouse; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases