Proteomic Analysis of Menstrual Blood

Abstract

Menstruation is the expulsion of the endometrial lining of the uterus following a nearly month long preparation for embryo implantation and pregnancy. Increasingly, the health of the endometrium is being recognized as a critical factor in female fertility, and proteomes and transcriptomes from endometrial biopsies at different stages of the menstrual cycle have been studied for both diagnostic and therapeutic purposes (1 Kao, L. C., et al. 2003 Endocrinology 144, 2870–2881; Strowitzki, Tet al. 2006 Hum. Reprod. Update 12, 617–630; DeSouza, L., et al. 2005 Proteomics 5, 270–281). Disorders of the uterus ranging from benign to malignant tumors, as well as endometriosis, can cause abnormal menstrual bleeding and are frequently diagnosed through endometrial biopsy (Strowitzki, Tet al. 2006 Hum. Reprod. Update 12, 617–630; Ferenczy, A. 2003 Maturitas 45, 1–14). Yet the proteome of menstrual blood, an easily available noninvasive source of endometrial tissue, has yet to be examined for possible causes or diagnoses of infertility or endometrial pathology. This study employed five different methods to define the menstrual blood proteome. A total of 1061 proteins were identified, 361 were found by at least two methods and 678 were identified by at least two peptides. When the menstrual blood proteome was compared with those of circulating blood (1774 proteins) and vaginal fluid (823 proteins), 385 proteins were found unique to menstrual blood. Gene ontology analysis and evaluation of these specific menstrual blood proteins identified pathways consistent with the processes of the normal endometrial cycle. Several of the proteins unique to menstrual blood suggest that extramedullary uterine hematopoiesis or parenchymal hemoglobin synthesis may be occurring in late endometrial tissue. The establishment of a normal menstrual blood proteome is necessary for the evaluation of its usefulness as a diagnostic tool for infertility and uterine pathologies. Identification of unique menstrual blood proteins should aid the forensic community in distinguishing menstrual blood from circulating blood. Menstruation is the expulsion of the endometrial lining of the uterus following a nearly month long preparation for embryo implantation and pregnancy. Increasingly, the health of the endometrium is being recognized as a critical factor in female fertility, and proteomes and transcriptomes from endometrial biopsies at different stages of the menstrual cycle have been studied for both diagnostic and therapeutic purposes (1 Kao, L. C., et al. 2003 Endocrinology 144, 2870–2881; Strowitzki, Tet al. 2006 Hum. Reprod. Update 12, 617–630; DeSouza, L., et al. 2005 Proteomics 5, 270–281). Disorders of the uterus ranging from benign to malignant tumors, as well as endometriosis, can cause abnormal menstrual bleeding and are frequently diagnosed through endometrial biopsy (Strowitzki, Tet al. 2006 Hum. Reprod. Update 12, 617–630; Ferenczy, A. 2003 Maturitas 45, 1–14). Yet the proteome of menstrual blood, an easily available noninvasive source of endometrial tissue, has yet to be examined for possible causes or diagnoses of infertility or endometrial pathology. This study employed five different methods to define the menstrual blood proteome. A total of 1061 proteins were identified, 361 were found by at least two methods and 678 were identified by at least two peptides. When the menstrual blood proteome was compared with those of circulating blood (1774 proteins) and vaginal fluid (823 proteins), 385 proteins were found unique to menstrual blood. Gene ontology analysis and evaluation of these specific menstrual blood proteins identified pathways consistent with the processes of the normal endometrial cycle. Several of the proteins unique to menstrual blood suggest that extramedullary uterine hematopoiesis or parenchymal hemoglobin synthesis may be occurring in late endometrial tissue. The establishment of a normal menstrual blood proteome is necessary for the evaluation of its usefulness as a diagnostic tool for infertility and uterine pathologies. Identification of unique menstrual blood proteins should aid the forensic community in distinguishing menstrual blood from circulating blood. Menstrual blood is a complex biological fluid composed of blood, vaginal secretions, and the endometrial cells of the uterine wall as they exist immediately prior to menses. These cells are the end product of a dynamic cyclical process focused on pregnancy and reproduction. Consequently, many of the proteins in these cells are expressed in preparation for blastocyst implantation and nurturing (2Strowitzki T. Germeyer A. Popovici R. von Wolff M. The human endometrium as a fertility-determining factor.Hum. Reprod. Update. 2006; 12: 617-630Crossref PubMed Scopus (210) Google Scholar, 5Garrido N. Navarro J. Garcia-Velasco J. Remoh J. Pellice A. Simón C. The endometrium versus embryonic quality in endometriosis-related infertility.Hum. Reprod. Update. 2002; 8: 95-103Crossref PubMed Scopus (145) Google Scholar). Other proteins in the menstrual blood proteome are a consequence of no implantation and include proteolytic enzymes, cytokines, members of apoptotic pathways, and a host of proteins from the diverse types of immune cells that are an integral part of menstruation (6Lommel A.T.L.V. From Cells to Organs. Kluwer Academic Publishers, Boston2003Crossref Google Scholar, 7King A.E. Critchley H.O. Oestrogen and progesterone regulation of inflammatory processes in the human endometrium.J. Steroid Biochem. Mol. Biol. 2010; 120: 116-126Crossref PubMed Scopus (96) Google Scholar, 8Ferenczy, A., Mutter, G, . (2008) The endometrial cycle. Glob. Libr. Women’s Med. 10.3843/GLOWM.10293Google Scholar, 9Li A. Felix J.C. Hao J. Minoo P. Jain J.K. Menstrual-like breakdown and apoptosis in human endometrial explants.Hum. Reprod. 2005; 20: 1709-1719Crossref PubMed Scopus (25) Google Scholar). Protein expression in all organs is a consequence of function. In the uterus, on a evaluation of the menstrual blood proteome on an of the complex uterine cycle. The uterus is composed of the the immediately and the a the The the in to the cycle and is is a the the and a the the menstrual cycle on the of and has an of at with the prior as the of the cycle and the following the the of the menstrual cycle and progesterone are at Menstruation the uterus is of the and of the (2Strowitzki T. Germeyer A. Popovici R. von Wolff M. The human endometrium as a fertility-determining factor.Hum. Reprod. Update. 2006; 12: 617-630Crossref PubMed Scopus (210) Google Scholar, 8Ferenczy, A., Mutter, G, . (2008) The endometrial cycle. Glob. Libr. Women’s Med. 10.3843/GLOWM.10293Google Scholar). as the endometrium is being from cells of the and as well as cells in the the process of the endometrial lining (6Lommel A.T.L.V. From Cells to Organs. Kluwer Academic Publishers, Boston2003Crossref Google Scholar, 8Ferenczy, A., Mutter, G, . (2008) The endometrial cycle. Glob. Libr. Women’s Med. 10.3843/GLOWM.10293Google Scholar). at the of the cycle and for a This is a of and and the of are in the of as well as the of and with in and and as progesterone in expression are as to (2Strowitzki T. Germeyer A. Popovici R. von Wolff M. The human endometrium as a fertility-determining factor.Hum. Reprod. Update. 2006; 12: 617-630Crossref PubMed Scopus (210) Google Scholar, L. J. analysis of the and of the human and 2005; PubMed Scopus Google Scholar). (6Lommel A.T.L.V. From Cells to Organs. Kluwer Academic Publishers, Boston2003Crossref Google Scholar, 8Ferenczy, A., Mutter, G, . (2008) The endometrial cycle. Glob. Libr. Women’s Med. 10.3843/GLOWM.10293Google the of in the and of at the of and the long that the the an found in the of the female endometrium (6Lommel A.T.L.V. From Cells to Organs. Kluwer Academic Publishers, Boston2003Crossref Google Scholar, 8Ferenczy, A., Mutter, G, . (2008) The endometrial cycle. Glob. Libr. Women’s Med. 10.3843/GLOWM.10293Google Scholar, T. J. and of the of normal human Google Scholar). The is necessary for the of synthesis by the endometrium in preparation for blastocyst implantation the of and of (6Lommel A.T.L.V. From Cells to Organs. Kluwer Academic Publishers, Boston2003Crossref Google Scholar, 8Ferenczy, A., Mutter, G, . (2008) The endometrial cycle. Glob. Libr. Women’s Med. 10.3843/GLOWM.10293Google Scholar). in by a of and (2Strowitzki T. Germeyer A. Popovici R. von Wolff M. The human endometrium as a fertility-determining factor.Hum. Reprod. Update. 2006; 12: 617-630Crossref PubMed Scopus (210) Google Scholar). as the late endometrial to preparation of of uterine and (2Strowitzki T. Germeyer A. Popovici R. von Wolff M. The human endometrium as a fertility-determining factor.Hum. Reprod. Update. 2006; 12: 617-630Crossref PubMed Scopus (210) Google Scholar, 8Ferenczy, A., Mutter, G, . (2008) The endometrial cycle. Glob. Libr. Women’s Med. 10.3843/GLOWM.10293Google Scholar). The endometrial lining in the and The of implantation no in of progesterone and to menses. of in apoptosis and A., Mutter, G, . (2008) The endometrial cycle. Glob. Libr. Women’s Med. 10.3843/GLOWM.10293Google Scholar, 9Li A. Felix J.C. Hao J. Minoo P. Jain J.K. Menstrual-like breakdown and apoptosis in human endometrial explants.Hum. Reprod. 2005; 20: 1709-1719Crossref PubMed Scopus (25) Google Scholar). the and with and inflammatory by the blood from and aid in the of the A., Mutter, G, . (2008) The endometrial cycle. Glob. Libr. Women’s Med. 10.3843/GLOWM.10293Google Scholar). The from the is the following as from cells and the of the uterus The health of the endometrium is a to pregnancy. or the to and a blastocyst can to have focused on the causes of endometrial and the for proteins that may be as diagnostic as well as therapeutic is Germeyer A. of endometrium from with for implantation and PubMed Scopus Google Scholar, T. Germeyer A. Popovici R. von Wolff M. The human endometrium as a fertility-determining factor.Hum. Reprod. Update. 2006; 12: 617-630Crossref PubMed Scopus (210) Google Scholar, L. J. analysis of the and of the human and 2005; PubMed Scopus Google Scholar). 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From Cells to Organs. Kluwer Academic Publishers, Boston2003Crossref Google Scholar, 8Ferenczy, A., Mutter, G, . (2008) The endometrial cycle. Glob. Libr. Women’s Med. 10.3843/GLOWM.10293Google Scholar, in proteins of human endometrial 2010; PubMed Scopus Google Scholar). endometrial in to immune cells and a embryo from its immune (6Lommel A.T.L.V. From Cells to Organs. Kluwer Academic Publishers, Boston2003Crossref Google Scholar, 8Ferenczy, A., Mutter, G, . (2008) The endometrial cycle. Glob. Libr. Women’s Med. 10.3843/GLOWM.10293Google Scholar). of implantation to menstruation is as an inflammatory by and uterine specific and through (2Strowitzki T. Germeyer A. Popovici R. von Wolff M. The human endometrium as a fertility-determining factor.Hum. Reprod. Update. 2006; 12: 617-630Crossref PubMed Scopus (210) Google Scholar, A.T.L.V. From Cells to Organs. Kluwer Academic Publishers, Boston2003Crossref Google Scholar, 7King A.E. Critchley H.O. Oestrogen and progesterone regulation of inflammatory processes in the human endometrium.J. Steroid Biochem. Mol. Biol. 2010; 120: 116-126Crossref PubMed Scopus (96) Google Scholar, 8Ferenczy, A., Mutter, G, . (2008) The endometrial cycle. Glob. Libr. Women’s Med. 10.3843/GLOWM.10293Google Scholar, in PubMed Scopus Google Scholar). 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Women’s Med. 10.3843/GLOWM.10293Google Scholar). has long been recognized that in the or of the endometrium can be diagnostic of uterine and is by abnormal (2Strowitzki T. Germeyer A. Popovici R. von Wolff M. The human endometrium as a fertility-determining factor.Hum. Reprod. Update. 2006; 12: 617-630Crossref PubMed Scopus (210) Google Scholar, 8Ferenczy, A., Mutter, G, . (2008) The endometrial cycle. Glob. Libr. Women’s Med. 10.3843/GLOWM.10293Google Scholar). for proteins have been with as well as endometrial In has as that can fertility, or infertility (2Strowitzki T. Germeyer A. Popovici R. von Wolff M. The human endometrium as a fertility-determining factor.Hum. Reprod. Update. 2006; 12: 617-630Crossref PubMed Scopus (210) Google Scholar). In the late as the uterus for the to with expression of or different of many of have been as of uterine proteins immune and A (2Strowitzki T. Germeyer A. Popovici R. von Wolff M. The human endometrium as a fertility-determining factor.Hum. Reprod. Update. 2006; 12: 617-630Crossref PubMed Scopus (210) Google Scholar, M. J.C. of endometrial preparation in a normal endometrial and in Reprod. 2006; PubMed Scopus Google Scholar). of these proteins were in the menstrual blood proteome and and may for is the proteome analysis of menstrual blood. The two were from cycle for as that the of a by can a menstrual blood proteome be with that of of and with or of is Menstrual blood is an easily fluid and is The that many of uterine infertility are in the menstrual blood proteome be for diagnostic menstruation can be a of uterine and a the normal menstrual blood proteome a diagnostic the of a of 385 menstrual blood proteins identified in blood or vaginal fluid an of for the menstrual blood in forensic to for of the with