PROSPECTIVE COMPARATIVE STUDY OF DIFFERENTIAL PROTEIN EXPRESSION IN TISSUE AND BLOOD SAMPLES OF MILD TO SEVERE FORMS OF ENDOMETRIOSIS UTILIZING PROTEOMICS

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This study used proteomics to identify seventeen differentially expressed proteins in blood and tissue samples of endometriosis patients compared to controls, proposing ZC3H13, SCUBE1, Afadin, and PRRC2A as potential biomarkers.

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This prospective comparative study used high-resolution two-dimensional gel electrophoresis combined with MALDI-TOF mass spectrometry to identify differential protein expression between tissue and fasting blood samples from 12 South Indian women undergoing laparoscopy for infertility, with six having endometriosis and six serving as controls without endometriosis. The authors reported that seventeen proteins were commonly expressed in both compartments of endometriosis patients versus controls and proposed a decision tree model using four named proteins (ZC3H13, SCUBE1, Afadin, and PRRC2A) to differentiate unexplained endometriosis with an accuracy of 0.97. The paper’s main limitation is the small sample size (six per group) and lack of an explicitly described independent validation cohort. This paper is centrally about endometriosis—using tissue and blood proteomics to compare mild-to-severe forms and propose candidate protein biomarkers.

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Abstract

Endometriosis is a complex condition characterized by cellular transformations in the body which plays a pivotal role in the pathogenesis and advancement of the condition. The integration of high-resolution two-dimensional gel electrophoresis (2-DE) analysis with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) facilitates the identification of proteins with differential expression in mild to severe forms of disease. Objective: This is the first study conducted in South Indian ethnic patients to compare the differential protein expression in mild to severe forms of endometriosis found in tissue and blood samples of the patients utilizing proteomics as its fundamental methodology. Methods: Twelve women with infertility as an issue have undergone laparoscopic procedures. The analysis included blood samples from six women diagnosed with EM (endometriosis) and six without EM during surgery, both groups having undergone biopsies. The blood sample was collected on the same day of surgery, at a fasting state, and was sent to the laboratory for further analysis of the protein expression in different groups i.e. Endometriosis and control group. Results: We identified seventeen proteins commonly expressed in blood and tissue samples of Endometriosis patients when compared with healthy controls and propose a panel of (A0PJJ2) methyltransferase zinc finger CCCH-type containing 13 (ZC3H13), Signal peptide, CUB, and EGF-like domain-containing proteins (SCUBE 1), Afadin (P55196), proline-rich coiled-coil 2A (PRRC2A) for differentiating women with unexplained Endometriosis from controls by using a decision tree model (accuracy: 0.97). Conclusion: The significantly upregulated proteins found in our study might pave the way in the field of endometriosis research further as potential biomarkers for early identification of the disease and for designing targeted and personalized treatment strategies. We propose that conducting diagnostic tests for both blood and tissue markers could enhance the precision of diagnosing Endometriosis.
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PROSPECTIVE COMPARATIVE STUDY OF DIFFERENTIAL PROTEIN EXPRESSION IN TISSUE AND BLOOD SAMPLES OF MILD TO SEVERE FORMS OF ENDOMETRIOSIS UTILIZING PROTEOMICS Authors/Creators - 1. MD (A.I.I.M.S, Delhi), F.R.C.O.G.(London), Professor and Head, Dept. of Obst & Gynecology, Shadan Institute of Medical Sciences, Medical and Research Director, Medical Health and Research Institute (MHRI), India. - 2. Pharm D, Research Scholar, Department of Obstetrics and Gynaecology, Medical Health and Research Institute (MHRI), India. - 3. Ph.D in Physiology, CMH Research and Innovations, Department of Cellular and Immunology, Bolarum, Secunderabad. - 4. M.Sc Genetics, Research Scholar, Department of Obstetrics and Gynaecology, Medical Health and Research Institute (MHRI), India. - 5. MD (Internal Medicine), Research Scholar, Department of Obstetrics and Gynaecology, Medical Health and Research Institute (MHRI), India. Description Endometriosis is a complex condition characterized by cellular transformations in the body which plays a pivotal role in the pathogenesis and advancement of the condition. The integration of high-resolution two-dimensional gel electrophoresis (2-DE) analysis with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) facilitates the identification of proteins with differential expression in mild to severe forms of disease. Objective: This is the first study conducted in South Indian ethnic patients to compare the differential protein expression in mild to severe forms of endometriosis found in tissue and blood samples of the patients utilizing proteomics as its fundamental methodology. Methods: Twelve women with infertility as an issue have undergone laparoscopic procedures. The analysis included blood samples from six women diagnosed with EM (endometriosis) and six without EM during surgery, both groups having undergone biopsies. The blood sample was collected on the same day of surgery, at a fasting state, and was sent to the laboratory for further analysis of the protein expression in different groups i.e. Endometriosis and control group. Results: We identified seventeen proteins commonly expressed in blood and tissue samples of Endometriosis patients when compared with healthy controls and propose a panel of (A0PJJ2) methyltransferase zinc finger CCCH-type containing 13 (ZC3H13), Signal peptide, CUB, and EGF-like domain-containing proteins (SCUBE 1), Afadin (P55196), proline-rich coiled-coil 2A (PRRC2A) for differentiating women with unexplained Endometriosis from controls by using a decision tree model (accuracy: 0.97). Conclusion: The significantly upregulated proteins found in our study might pave the way in the field of endometriosis research further as potential biomarkers for early identification of the disease and for designing targeted and personalized treatment strategies. We propose that conducting diagnostic tests for both blood and tissue markers could enhance the precision of diagnosing Endometriosis. Files 3 WJPSR 301.pdf Files (945.1 kB) | Name | Size | Download all | |---|---|---| | md5:e7b13318fd8bc6bdc27eae22ea12cf02 | 945.1 kB | Preview Download | Additional details References - Smolarz, B., Szyłło, K., & Romanowicz, H., Endometriosis: Epidemiology, Classification, Pathogenesis, Treatment and Genetics (Review of Literature). International journal of molecular sciences, 2021; 22(19): 10554. https://doi.org/10.3390/ijms221910554 - Klemmt, P. A. B., & Starzinski-Powitz, A., Molecular and Cellular Pathogenesis of Endometriosis. Current women's health reviews, 2018; 14(2): 106–116. https://doi.org/10.2174/1573404813666170306163448 - Macer, M. L., & Taylor, H. S., Endometriosis and infertility: a review of the pathogenesis and treatment of endometriosis-associated infertility. Obstetrics and gynecology clinics of North America, 2012; 39(4): 535–549. https://doi.org/10.1016/j.ogc.2012.10.002 - Zhang, H., Niu, Y., Feng, J., Guo, H., Ye, X., & Cui, H., Use of proteomic analysis of endometriosis to identify different protein expression in patients with endometriosis versus normal controls. Fertility and sterility, 2006; 86(2): 274–282. https://doi.org/10.1016/j.fertnstert.2006.01.028 - Nisenblat, V., Bossuyt, P. M., Shaikh, R., Farquhar, C., Jordan, V., Scheffers, C. S., Mol, B. W., Johnson, N., & Hull, M. L., Blood biomarkers for the non-invasive diagnosis of endometriosis. The Cochrane database of systematic reviews, 2016; 2016(5): CD012179. https://doi.org/10.1002/14651858.CD012179 - Anastasiu, C. V., Moga, M. A., Elena Neculau, A., Bălan, A., Scârneciu, I., Dragomir, R. M., Dull, A. M., & Chicea, L. M., Biomarkers for the Noninvasive Diagnosis of Endometriosis: State of the Art and Future Perspectives. International journal of molecular sciences, 2020; 21(5): 1750. https://doi.org/10.3390/ijms21051750 - Laudanski, P., Charkiewicz, R., Tolwinska, A., Szamatowicz, J., Charkiewicz, A., & Niklinski, J., Profiling of Selected MicroRNAs in Proliferative Eutopic Endometrium of Women with Ovarian Endometriosis. BioMed research international, 2015; 2015: 760698. https://doi.org/10.1155/2015/760698 - Cosar, E., Mamillapalli, R., Ersoy, G. S., Cho, S., Seifer, B., & Taylor, H. S., Serum microRNAs as diagnostic markers of endometriosis: a comprehensive array-based analysis. Fertility and sterility, 2016; 106(2): 402-409. - Haikalis, M. E., Wessels, J. M., Leyland, N. A., Agarwal, S. K., & Foster, W. G., MicroRNA expression pattern differs depending on endometriosis lesion type. Biology of reproduction, 2018; 98(5): 623-633. - Rozati, R., Khan, A. A., Mehdi, A. G., Tabasum, W., & Begum, S., Circulating oncomiR-21 as a potential biomarker of mild and severe forms of endometriosis for early diagnosis and management. Gene Reports, 2023; 30: 101725. - Rozati, R., Mohiuddin, N., & Ayapati, V. A., Differential Protein Expression of mild to severe forms of Endometriosis in South Indian Ethnic patients. Gynecol Reprod Health, Correspondence: Roya Rozati, Medical and Research Director, Maternal Health and Research Trust, Banjara Hills, Hyderabad, India. Received, 2023; 7(5): 1-11. - Liu, A. X., Jin, F., Zhang, W. W., Zhou, T. H., Zhou, C. Y., Yao, W. M., Qian, Y. L., & Huang, H. F., Proteomic analysis on the alteration of protein expression in the placental villous tissue of early pregnancy loss. Biology of reproduction, 2006; 75(3): 414–420. https://doi.org/10.1095/biolreprod.105.049379 - Bahramy, A., Zafari, N., Izadi, P., Soleymani, F., Kavousi, S., & Noruzinia, M., The Role of miRNAs 340-5p, 92a-3p, and 381-3p in Patients with Endometriosis: A Plasma and Mesenchymal Stem-Like Cell Study. BioMed research international, 2021; 2021: 5298006. https://doi.org/10.1155/2021/5298006 - ZC3H13 zinc finger CCCH-type containing 13 [Homo sapiens (human)] - Gene - NCBI. (n.d.). https://www.ncbi.nlm.nih.gov/gene?Db=gene&Cmd=DetailsSearch&Term=23091 - Gu, J., Zhan, Y., Zhuo, L., Zhang, Q., Li, G., Li, Q., Qi, S., Zhu, J., Lv, Q., Shen, Y., Guo, Y., Liu, S., Xie, T., & Sui, X., Biological functions of m6A methyltransferases. Cell & bioscience, 2021; 11(1): 15. https://doi.org/10.1186/s13578-020-00513-0 - Nadezhdin, K.D., Correia, L., Narangoda, C. et al. Structural mechanisms of TRPM7 activation and inhibition. Nat Commun, 2023; 14: 2639. https://doi.org/10.1038/s41467-023-38362-3 - Takai, Y., & Nakanishi, H., Nectin and afadin: novel organizers of intercellular junctions. Journal of cell science, 2003; 116(Pt 1): 17–27. https://doi.org/10.1242/jcs.00167 - Ikeda, W., Nakanishi, H., Miyoshi, J., Mandai, K., Ishizaki, H., Tanaka, M., Togawa, A., Takahashi, K., Nishioka, H., Yoshida, H., Mizoguchi, A., Nishikawa, S., & Takai, Y., Afadin: A key molecule essential for structural organization of cell-cell junctions of polarized epithelia during embryogenesis. The Journal of cell biology, 1999; 146(5): 1117–1132. https://doi.org/10.1083/jcb.146.5.1117 - Wu, R., Li, A., Sun, B., Sun, J. G., Zhang, J., Zhang, T., Chen, Y., Xiao, Y., Gao, Y., Zhang, Q., Ma, J., Yang, X., Liao, Y., Lai, W. Y., Qi, X., Wang, S., Shu, Y., Wang, H. L., Wang, F., Yang, Y. G., … Yuan, Z., A novel m6A reader Prrc2a controls oligodendroglial specification and myelination. Cell research, 2019; 29(1): 23–41. https://doi.org/10.1038/s41422-018-0113-8. - Kumar, S., Prajapati, K. S., & Gupta, S., The Multifaceted Role of Signal Peptide-CUB-EGF Domain-Containing Protein (SCUBE) in Cancer. International journal of molecular sciences, 2022; 23(18): 10577. https://doi.org/10.3390/ijms231810577.

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