Higher order of endometriosis gene-expression, chromosome 1q significantly contributes to endometriosis specific gene-expression, meta-analysis of three (3)independent endometriosis studies

1. Gene-expression in Endometriosis: Chromosome 1q significantly contributes to endometriosis gene-expression MehisPold, M.D. Founder EOMIX, Inc. August 9, 2010 8/9/2010 1 mehis.pold@eomix.com

2. Endometriosis (from endo, "inside", and metra, "womb") is a medical condition in which endometrial cells are deposited in areas outside the uterine cavity. The uterine cavity is lined by endometrial cells, which are under the influence of female hormones. Endometrial cells deposited in areas outside the uterus (endometriosis) continue to be influenced by these hormonal changes and respond similarly as do those cells found inside the uterus. Symptoms often exacerbate in time with the menstrual cycle. Endometriosis is typically seen during the reproductive years; it has been estimated that it occurs in roughly 5% to 10% of women. Symptoms depend on the site of implantation. Its main but not universal symptom is pelvic pain in various manifestations. Endometriosis is a common finding in women with infertility. Source: Wikipedia Terms: Ectoendometrium – endometrial tissue outside of it’s natural location Euendometrium – endometrium in it’s natural location, uterine cavity 8/9/2010 2 mehis.pold@eomix.com

3. Diagnostics of Endometriosis: INVASIVE - Confirmed by laparoscopy Exclusion of other conditions Systemic diagnostic markers poorly understood Medical need for non-invasive diagnostic test How do we find endometriosis markers? Blood Urine Saliva Skin biopsy Hair follicles 8/9/2010 3 mehis.pold@eomix.com

4. (1) Meta-analysis of three independent whole-genome expression sets (2) GenMAPP pathway analysis Endometriosis database of EOMIX Inc. 8/9/2010 4 mehis.pold@eomix.com

5. Summary Statistics of Endometriosis sets (Roth, Klinkova, Hull) Pathway analysis by GenMAPP on genes altered in at least two of out three Endometriosis datasets N = 2,522 = Venn diagram [2+3+4] R = 19,632 8/9/2010 5 mehis.pold@eomix.com

8. GenMAPP Overview Small amount of information does not produce reliable results Results depend on Gene Ontology hierarchies and the existing pathway information Z-values greater than 2 and smaller than -2 considered relevant Z < -2 and Z > 2 Examples at R/N= 0.128 (12.8%) n = 90 pathway of 90 known members, r = 30 of which 30 are altered r/n = 0.333 (33.3%) Z = 5.82 n = 9 r = 3 r/n = 0.333 (33.3%) Z = 1.83 = insignificant 8/9/2010 7 mehis.pold@eomix.com

10. Actin filament-based process

11. Anion transport

12. Cell growth

13. Cell-cell adhesion

14. Endocytosis

15. Enzyme-linked receptor protein signaling

16. Growth

17. Humoral Defense sensu Vertebrata

18. Humoral Immune response

19. I-kappa kinase NF-kappa cascade

20. Inflammatory response

21. Inorganic anion transport

22. Muscle contraction

23. Muscle development

24. Regulation of cell size

26. Cytoskeleton

27. Extracellular matrix sensuMetazoa

29. Carbohydrate binding

30. Enzyme activator activity

31. Enzyme inhibiting activity

33. G13 signalin

34. G-protein signaling

35. Integrin-mediated adhesion

37. Calcium regulation in cardiac muscle

38. Circadian exercise

39. Complement activation by classical pathway

40. Complement and coagulation cascade KEGG

41. Smooth muscle contraction

42. Striated muscle contraction8/9/2010 8 mehis.pold@eomix.com

44. Glutathione metabolism

45. Lysine biosynhesis

46. Lysine degradation

47. Methionine metabolism

48. Nicotinate and nicotinamide metabolism

49. One carbon pool by folate

50. Phe, Tyr, Trp biosynthesis

51. Selenoamina acid metabolism

53. Cytokinesis

54. DNA repair

55. DNA replication

56. M phase

57. M phase of mitotic cell cycle

58. Microtubule-based process

59. Regulation of cell cycle

60. Response to DNA damage stimuli

62. Endomembrane system

64. Nucleotidyltrasferase activity

66. Cell cycle G1 to S control

67. Delta-Notch signaling

68. DNA replication reactome

69. Hedgehog pathway

70. mRNA processing reactome

72. Fatty acid synthesis

74. Mismatch repairPhysiological Process 8/9/2010 9 mehis.pold@eomix.com

76. Muscle-related pathways upregulated in endometriosis: smooth and striated muscle-related genes, calcium-regulated gene-expression.

77. Cytoskeleton-related processes involved

78. Metabolism and Cell cycle appear low in endometriosis as compared to normal endometrial tissueConnection to previously published studies Plasma cell infiltration ~ high levels of B-cell receptor gene-expression documented in previous studies. Heveret al. (PNAS, 2007) documented plasma-cell infiltration of ovarian endometriosis. 8/9/2010 10 mehis.pold@eomix.com

79. Chromosome 1q produces substantial levels of endometriosis-specific gene-expression 8/9/2010 11 mehis.pold@eomix.com

80. Chromosome 1q32 genes that exhibit altered expression in endometriosis Red font and blue font denote up- and downregulation, respectively 8/9/2010 12 mehis.pold@eomix.com

81. SUMMARY Pathways modulating host defense are activated in endometriosis Chromosome 1q significantly contributes to endometriosis gene-expression 8/9/2010 mehis.pold@eomix.com 13