Deryabin Pavel, Domnina Alisa, Gorelova Inga, Rulev Maxim, Petrosyan Mariya, Nikolsky Nikolay, Borodkina Aleksandra
Mechanisms of Cellular Senescence Group, Institute of Cytology of the Russian Academy of Sciences, Saint-Petersburg, Russia.
Department of Intracellular Signaling and Transport, Institute of Cytology of the Russian Academy of Sciences, Saint-Petersburg, Russia.
Front Cell Dev Biol. 2021 Feb 15;9:624053. doi: 10.3389/fcell.2021.624053. eCollection 2021.
Endometrium is the uterine lining that undergoes hundreds of cycles of proliferation, differentiation, and desquamation throughout a woman's reproductive life. Recently, much attention is paid to the appropriate endometrial functioning, as decreased endometrial receptivity is stated to be one of the concerns heavily influencing successes of embryo implantation rates and the efficacy of fertilization (IVF) treatment. In order to acquire and maintain the desired endometrial receptivity during IVF cycles, luteal phase support by various progestagens or other hormonal combinations is generally recommended. However, today, the selection of the specific hormonal therapy during IVF seems to be empirical, mainly due to a lack of appropriate tools for personalized approach. Here, we designed the genetic tool for patient-specific optimization of hormonal supplementation schemes required for the maintenance of endometrial receptivity during luteal phase. We optimized and characterized endometrial stromal cell (ESC) decidualization model as the adequate physiological reflection of endometrial sensitivity to steroid hormones. Based on the whole transcriptome RNA sequencing and the corresponding bioinformatics, we proposed that activation of the decidual prolactin (PRL) promoter containing ancient transposons MER20 and MER39 may reflect functioning of the core decidual regulatory network. Furthermore, we cloned the sequence of decidual PRL promoter containing MER20 and part of MER39 into the expression vector to estimate the effectiveness of ESC decidual response and verified sensitivity of the designed system. We additionally confirmed specificity of the generated tool using human diploid fibroblasts and adipose-derived human mesenchymal stem cells. Finally, we demonstrated the possibility to apply our tool for personalized hormone screening by comparing the effects of natural progesterone and three synthetic analogs (medroxyprogesterone 17-acetate, 17α-hydroxyprogesterone caproate, dydrogesterone) on decidualization of six ESC lines obtained from patients planning to undergo the IVF procedure. To sum up, we developed the "all-in-one" genetic tool based on the MER20/MER39 expression cassette that provides the ability to predict the most appropriate hormonal cocktail for endometrial receptivity maintenance specifically and safely for the patient, and thus to define the personal treatment strategy prior to the IVF procedure.
子宫内膜是子宫的内衬，在女性的生殖生活中会经历数百次增殖、分化和脱屑循环。最近，人们非常关注子宫内膜的正常功能，因为子宫内膜容受性降低被认为是严重影响胚胎着床率和体外受精（IVF）治疗效果的问题之一。为了在IVF周期中获得并维持理想的子宫内膜容受性，通常建议使用各种孕激素或其他激素组合进行黄体期支持。然而，如今在IVF期间选择特定的激素疗法似乎是经验性的，主要是因为缺乏用于个性化治疗的合适工具。在此，我们设计了一种基因工具，用于针对患者特异性优化黄体期维持子宫内膜容受性所需的激素补充方案。我们优化并表征了子宫内膜基质细胞（ESC）蜕膜化模型，将其作为子宫内膜对类固醇激素敏感性的适当生理反映。基于全转录组RNA测序和相应的生物信息学，我们提出含有古老转座子MER20和MER39的蜕膜催乳素（PRL）启动子的激活可能反映核心蜕膜调节网络的功能。此外，我们将含有MER20和部分MER39的蜕膜PRL启动子序列克隆到表达载体中，以评估ESC蜕膜反应的有效性，并验证所设计系统的敏感性。我们还使用人二倍体成纤维细胞和脂肪来源的人间充质干细胞证实了所生成工具的特异性。最后，我们通过比较天然孕酮和三种合成类似物（醋酸甲羟孕酮、己酸羟孕酮、地屈孕酮）对从计划接受IVF手术的患者获得的六个ESC系蜕膜化的影响，证明了应用我们的工具进行个性化激素筛选的可能性。总之，我们基于MER20/MER39表达盒开发了一种“一体化”基因工具，该工具能够为患者特异性且安全地预测维持子宫内膜容受性的最合适激素组合，从而在IVF手术前确定个性化治疗策略。
