Dataset Information

Proteomics-Based Retinal Target Engagement Analysis and Retina-Targeted Delivery of 17β-Estradiol by the DHED Prodrug in Males. Katalin Prokai-Tatrai,* Khadiza Zaman, Vien Nguyen, Daniel L. De La Cruz and Laszlo Prokai

ABSTRACT: We examined the impact of 17β-estradiol (E2) eye drops on the modulation of the proteome pro-file in the male rat retina. With discovery-driven proteomics, we have identified proteins that were regulated by the treatment. These proteins were assembled to several bioinformatics-based networks implicating E2’s beneficial effects on the male rat retina in a broad context of ocular neuroprotection including the maintenance of retinal homeostasis, facilitation of efficient dis-posal of damaged proteins, and mitochondrial respiratory chain biogenesis. We have also shown for the first time that the hormone’s beneficial effects on the male retina can be con-strained to this target site by treatment with the bioprecursor prodrug DHED. A large concen-tration of E2 was produced after DHED eye drops not only in male rat retinae but also in those of rabbits. However, DHED treatment did not increase circulating E2 levels thereby ensuring therapeutic safety in males. Targeted proteomics focusing on selected biomarkers of E2’s target engagement further confirmed the prodrug’s metabolism to E2 in the male retina and indicated that the retinal impact of DHED treatment was identical to that of the direct E2 treatment. Alto-gether, our study shows the potential of topical DHED therapy for an efficacious and safe protec-tion of the male retina without unwanted hormonal side-effects associated with current estrogen therapies.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Rattus Norvegicus (rat)

TISSUE(S): Retina

SUBMITTER: Laszlo Prokai

LAB HEAD: Laszlo Prokai, Ph.D

PROVIDER: PXD027902 | Pride | 2021-11-04

REPOSITORIES: Pride

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Publications

Proteomics-Based Retinal Target Engagement Analysis and Retina-Targeted Delivery of 17β-Estradiol by the DHED Prodrug for Ocular Neurotherapy in Males.

Prokai-Tatrai Katalin K Zaman Khadiza K Nguyen Vien V De La Cruz Daniel L DL Prokai Laszlo L

Pharmaceutics 20210902 9

We examined the impact of 17β-estradiol (E2) eye drops on the modulation of the proteome profile in the male rat retina. With discovery-driven proteomics, we have identified proteins that were regulated by our treatment. These proteins were assembled to several bioinformatics-based networks implicating E2's beneficial effects on the male rat retina in a broad context of ocular neuroprotection including the maintenance of retinal homeostasis, facilitation of efficient disposal of damaged proteins ...[more]

PMID: 34575465

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Project description:E2 and GH are critical regulators of growth and intermediate metabolism in mammals. Hypothyroidism causes endocrine and metabolic disturbances in the liver with features that mimic deficiencies of E2 or GH signalling. In this work, we used the hypothyroid-orchiectomized (TXOX) adult rat model to evaluate the influence of E2 and GH on the liver in terms of global changes in gene expression. This study shows the changes in hepatic transcriptome that were provoked by E2 benzoate (50 ug/kg; sc; 5 days per week x 27 days), intermittent GH administration (0.3 mg/kg/day;sc injection divided into two daily injections x 7 days) or the combination of E2 plus GH in TXOX rats. E2 influenced the liver transcriptome, particularly genes involved in metabolism of lipids and endo-xenobiotics, and the GH-regulated endocrine, metabolic, gender, and immune responses. E2 did not prevent the inhibitory effects of GH on urea and amino acid metabolism-related genes. Notably, the combination of E2 and GH caused deleterious effects on transcriptional immune response. These results highlight the role of E2 as a critical regulator of liver metabolism in mammals and provide insights into the functional interplay between E2 and GH in the liver. Groups=4; Biological replicates = 4 per group; Samples=16; Reference samples=TXOX group. Adult (3 months old) male Sprague-Dawley rats (n=4 per group) were used throughout these experiments. The generation of TXOX was performed by adding methimazole (MMI; 0.05%) to the drinking water for 5 weeks starting on postnatal day (PND) 59 until sacrifice on PND94. Two weeks after starting MMI administration, male rats were castrated (OX) to make TXOX rats. Four days after OX, we began treatment with E2 benzoate (TXOXE2) or vehicle (TXOX) to TXOX rats for 20 days followed for 7 days by either vehicle plus GH (TXOXGH) or by E2 plus GH (TXOXE2GH). Twenty-four hours (in the case of E2) or twelve hours (in the case of GH) after the last injection, the animals were killed by exsanguinations. Portions of the liver were snap frozen in liquid nitrogen and stored at -80C until processed for mRNA analysis.

Project description:Corpus luteum (CL) is a transient endocrine tissue formed from the remnants of the ovarian follicle after ovulation. In response to gonadotropin surge, the ovulating follicle undergoes dramatic changes in expression of genes and differentiation of follicular cells into luteal cells. In several species, of the several genes that are down- regulated post ovulation, Cyp19A1 that codes for aromatase, essential for biosynthesis of estradiol- 17M-NM-2 (E2), also get down- regulated but appears to get up- regulated at later time points in the estrous cycle to have critical role in E2 secretion. In primates and rodents, higher expression and higher E2 levels has been observed in CL. Surprisingly, in the recently carried out gene expression profiling of PGF2M-NM-1- induced luteolysis studies in the bovine species [GSE27961], it was observed that expression of one of the earliest genes that was down- regulated was Cyp19A1 in the CL. However, the specific role of E2 in the regulation of CL function remains poorly defined. Thus, in the present study, efforts were made to examine the temporal changes in the global gene expression profile in the CL of pregnant rats after treatment with aromatase inhibitor (AI), Anastrozole, and E2 supplementation. The results obtained will further expand our knowledge on E2 target/responsive genes and the basic mechanism(s) that regulates the CL function. Key words: CL, E2, AI, Gene expression The objective of this study was to identify the effect of E2 deprivation and supplementation on CL function and temporal changes in the transcriptome of pregnant rat CL during day 12 to 16 of pregnancy following E2 deprivation by Anastrozole (AI, Aromatase inhibitor) treatment in pregnant rats, AI (1mg/ kg bw orally) treatment beginning on day 12 daily for 4 days. In the E2 supplementation experiments, together with AI, the rats were treated with E2 (10M-BM-5g/ day s.c.). CL tissues were collected on day 12 (no treatment; n=3 animals/ time point), day 16 (vehicle treatment; n=3 animals/ time point), day 16 (AI treatment; n=3 animals/ time point) and day 16 (AI+ E2 treatment; n=3 animals/ time point) to examine gene expression changes associated with E2 deprivation and recovery in the system. Following retrieval of CL from the ovary, corpora lutea were flash frozen in liquid nitrogen and stored at -70oC for the purpose of RNA isolation. The isolated RNA was labelled and hybridized to Affymetrix GeneChipM-BM-. Rat Gene 1.0 ST arrays as per the manufacturerM-bM-^@M-^Ys instructions.

Dataset Information

Proteomics-Based Retinal Target Engagement Analysis and Retina-Targeted Delivery of 17β-Estradiol by the DHED Prodrug in Males. Katalin Prokai-Tatrai,* Khadiza Zaman, Vien Nguyen, Daniel L. De La Cruz and Laszlo Prokai

Publications

Proteomics-Based Retinal Target Engagement Analysis and Retina-Targeted Delivery of 17β-Estradiol by the DHED Prodrug for Ocular Neurotherapy in Males.

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