Cross-talk of the microbial Transcriptional response to Polycystic Ovary Syndrome alteration modulate of mouse gut

The presentation shows an example to create ideas from scientific papers using microbiome approaches with different topics. I was using these approaches to figure out the gaps in the specific topics in this case Polycystic Ovary Syndrome ( PCOS) after reviewing the literature as an example and presented to the advisor. These methods are a helpful example to explain how to get an idea and organize any new research topics in particular the new graduate students.

Transcript

1. SUAD ALGARNI PROPOSAL COMMITTEE MEETING CELL AND MOLECULAR BIOLOGY PROGRAM, ANIMAL SCINCES DEPARTMENT Cross-talk of the microbial transcriptional response to Polycystic Ovary syndrome alteration modulate of mouse model gut microbiome
2. Outline • Introduction • Background • Objective • Approach • Future work
3. Introduction • Polycystic Ovary Syndrome (PCOS) is endocrine disorder characterized by irregular menses, hyperandrogenism, and polycytic ovaries. • PCOS affects 4%-8% of women of reproductive age. • Prevalence of 15–20% based on the European Society for Human Reproduction and Embryology/American Society for Reproductive Medicine creiteria. • In worldwide, PCOS affecting 5%-10% of women.
4. Background • Obesity increased the risk of PCOS. • 40.5% of women PCOS in the type 1 diabetes. • 82% of women PCOS in type 2 diabetes. • 30%-40% of women with PCOS have glucose tolerance. • PCOS cause mental health disorders: (Depression, anxiety, bipolar disorder and binge eating disorder) • Cardiovascular disease • hypertension E Carmina et al. Eur J Endocrinol 2006;154:141-145 Prevalence of different elements of the metabolic syndrome in women with PCOS
5. Diagnostic Criteria for PCOS:
6. Signs & Symptoms of PCOS • Hirsutism • Infertility • Obesity and metabolic syndrome • Diabetes • Obstructive sleep apnea (OSA)
7. Treatment • NO SPECIFIC TREATMENT FOR PCOS Treatment of PCOS depends on the symptoms: 1. Obesity (e.g. Metformin) 2. Excess hair ( e.g. vaniqa, laser) 3. Fertility (e.g. Letrozole, clomid) • Lifestyle change ( diet and exercise)
8. Dysbiosis of gut microbiota (DOGMA) by a high fat-sugar diet • Gut microbiota in PCOS, as the androgen level in PCOS women always increased. • Lipopolysaccharide produced by gram negative bacteria lead to low grade of inflammation •The activation of immune system interferes with insulin receptor, driving up insulin level, which boosts testosterone production in the ovary leading to PCOS. Guo Y, Qi Y, Yang X, Zhao L, Wen S, et al. (2016) Association between Polycystic Ovary Syndrome and Gut Microbiota. PLOS ONE 11(4): e0153196. doi:10.1371/journal.pone.0153196
9. PCOS mouse model • Letrozole ( a non steroidal aromatase inhibitor) treatment induced PCOS mouse model. Concentration (50 ug/day) 5 weeks= 35 days • Changes in estrous cycles • Hormonal levels • Ovarian morphology • Gut microbiots • Body weight Guo, Y. et al. 2016. Association between Polycystic Ovary Syndrome and Gut Microbiota. PLOS ONE. 11, 4 (2016), e0153196
10. Letrozole treatment of 4-week-old female mice resulted in both reproductive and metabolic hallmarks of PCOS Kelley ST, Skarra DV, Rivera AJ, Thackray VG (2016) The Gut Microbiome Is Altered in a Letrozole- Induced Mouse Model of Polycystic Ovary Syndrome. PLOS ONE 11(1): e0146509. doi:10.1371/journal.pone.0146509 http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0146509 Increased serum testosterone Polycystic ovaries Increased body weight Increased fat mass
11. Did the Letrozole treatment alter the gut of PCOS mice model? Kelley ST, Skarra DV, Rivera AJ, Thackray VG (2016) The Gut Microbiome Is Altered in a Letrozole-Induced Mouse Model of Polycystic Ovary Syndrome. PLOS ONE 11(1): e0146509. doi:10.1371/journal.pone.0146509 http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0146509 Ø Associated with a time dependent shift in the gut microbiome. Ø Correlated with significant changes in the abundance of specific Bacteroidetes and Firmicutes in other mouse models of metabolic disease.
12. Question • Does the PCOS an ovarian or metabolic disorder disease? • What is the role functional effect of altered gut microbiome of PCOS? • What is the absolute abundance of specific bacterial species effect of letrozole on gut microbiome?
13. Overall goal • Understand the role pathogenesis of gut microbime of PCOS. • Investigate the associated between metabolic pathway and gene identification of PCOS. • Study the primary pathophysiology mechanism of PCOS.
14. Approach • 16S rRNA gene microbiome • Metagenomic • Metatranscriptomics
15. Hypothesis • We hypothesize that diet manipulation will shape the gut microbiome of PCOS with letrozole (LET).
16. Objective • Investigate the effect composition of gut microbiome by using low carbs and fiber rich diet with letrozole treatment.
17. Approach • Experimental design: • Period time: • Pre PCOS • Post PCOS • After PCOS n= 4 Control n= 5 mice High fiber diet(HFD) + LETROZLE n= 5 mice low carbs diet(HFD) + LETROZLE
18. Characterize the gut microbial communities: Collect fecal sample Extract DNA quantify quality PCR Amplify + library preparation V4 hypervariable region Cluster OTU
19. Next step • Objective 1: Ø Perform sample processing for animal experimental Ø Extract DNA from fecal samples Ø Illumina sequencing and data analysis