MSACL 2016 EU Abstract

Endometriotic Tissues Differentiation by Mass Spectrometry

Vladimir Frankevich (Presenter)
Research Center for obstetrics and gynecology

Bio: Head of Systems Biology Department, PhD

Authorship: Vladimir Frankevich, Vitaly Chagovets, Ànna. Borisova, Natalia Starodubtseva, Àlexey Kononikhin, Leila Adamyan
Scientific Center for Obstetrics, Gynecology and Perinatology of the Ministry of Health of the Russian Federation.Moscow, Russia

Short Abstract

Development of instant and non-invasive endometriotic tissues differentiation method. For the first time the Direct Mass Spectrometry approach was applied to analyze endometrium and endometriotic lesions of different localization. Our clinical lipidomics approach defines the net outcome of several imbalanced lipids and fatty acids which may be crucial for endometriosis pathophysiology and tissues differentiation. The results of these studies may bring us closer to the understanding of the pathobiology of the disease and make it possible to elaborate new ways of early diagnostics and “intelligent” surgery.

Long Abstract

Introduction

Endometriosis is a gynecological disorder characterized by presence of endometrial tissue outside the uterine cavity. The pathogenesis of endometriosis is still unknown and current therapies are only symptomatic. The main method of treatment is complete removal of endometriosis foci by surgical methods. An important task is the quality control of removal of the endometriosis foci and prediction of disease relapses. It is not so easy for surgeon to visualize the border of the healthy tissue. Mass spectrometry (MS) is the method of choice for this purpose.

MS is one of the most widely used techniques for the analysis of biological samples. In the past decade, a novel improvement in MS was the invention of ambient ionization (direct MS) which stands out owing to its unique capability of direct analysis of complex samples with no or minimal pretreatment.

The aim of this research was the development of fast MS method for analysis of endometriotic and other tissues. This method will be a key instrument for the “intelligent” gynecological surgery.

Methods

All the samples were collected during surgical procedures in the department of Operative Gynecology of the Scientific Center of obstetrics, gynecology and perinatology (Moscow, Russia). On the case–control study we recruited 80 patients with ovarian cysts and with peritoneal endometriosis who then underwent laparoscopic surgery between 2015 and 2016. The diagnosis of endometriosis was confirmed histologically. According to this classification, 40 patients had I-II stage of endometriosis, the other 40 patients - III-IV stage. The primary inclusion criteria were: reproductive age, absence of any severe somatic pathologies, inflammatory diseases.The endometriotic lesions after excision (without thermal effect), the capsules of endometrioid cyst after cystectomy, the endometrium after curettage were immediately placed in liquid nitrogen, followed by freeze-up at –800 Ñ. Differences in the mass spectrometric profiles of the endometriotic tissues and comparison group were analyzed in combination with the morphological features in order to find the specific biomarkers.

Direct-spray-from-tissue method was used for on-line molecular species extraction and simultaneously ionization. All MS and MS2 spectra were acquired by electrospray ionization quadrupole time of flight mass spectrometer (Maxis Impact, Bruker, Germany).

Results

148 metabolites were analyzed. Phosphoethanolamine (PE O-20:0), sphingomyelin (SM 34:1), diglycerides (DG 44:9), phosphatidylcholines (PC 32:1, PC O-36:3, PC 38:7, PC 38:6, PC 40:8, PC 40:7, PC 40:6, PC O-42:1), triglycerides (TG 41:2, TG 49:4, TG 52:3) showed significant differences between the biological samples of the patients with endometriosis and those of the control group. The procedure of endometriotic tissues differentiation was developed for further implementation into “intelligent” surgery.

Conclusion

For the first time the Direct MS approach was applied to analyze endometrium and endometriotic lesions of different localization. Our clinical lipidomics approach defines the net outcome of several imbalanced lipids and fatty acids which may be crucial for endometriosis pathophysiology and tissues differentiation. Two important classes of lipids, the phosphatidylcholine and sphingomyelin, have been suggested as possible biomarkers for endometriosis. The results of these studies may bring us closer to the understanding of the pathobiology of the disease and make it possible to elaborate new ways of early diagnostics and “intelligent” surgery.


References & Acknowledgements:

This work was supported by grant No. 16-14-00029 of RSF.


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