Assisted Reproductive. Technologies: Present and. Future
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1 Assisted Reproductive Technologies: Present and Future Paul Kaplan, M.D.
2 The Assisted Reproductive Technologies (ART) In Vitro Fertilization (IVF) Intracytoplasmic Sperm Injection (IVF/ICSI) Donor Oocyte IVF Frozen Embryo Thaw and Transfer Cryopreservation/In Vitro Maturation of Oocytes
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4 In Vitro Fertilization (IVF) Daily S/C or IM FSH/hMG injection Follicular monitoring with serum estradiol and transvaginal ultrasound HCG given to trigger ovulation (LH surge) Transvaginal oocyte retrieval and insemination Embryo culture and transcervical embryo transfer Embryo cryopreservation for future F.E.T. Pregnancy rate of % per cycle
5 IVF Embryo Culture and Transfer
6 Intracytoplasmic Sperm Injection (ICSI) Standard IVF Stimulation and oocyte retrieval Injection of a single sperm into each oocyte Embryo culture and transcervical embryo transfer Currently used in almost 50% of IVF cycles for treatment of male factor and unexplained causes Pregnancy rate of % per cycle
7 Intracytoplasmic Sperm Injection (ICSI)
8 Future Directions in ART The omics revolution in non-invasive screening Preimplantation genetic diagnosis (PGD) - with gene therapy? Nuclear and/or cytoplasmic oocyte transfer Embryonic Stem Cell Line Development Gamete Stem Cell Development
9 Future Directions in ART (Con t) Embryo Cloning - Reproductive/Therapeutic Adult Cell Gamete Cloning - sperm/oocyte Adult Somatic Cell Cloning
10 The -omics Revolution in Infertility Genomics: The branch of molecular biology concerned with the structure, function, evolution, and mapping of genomes. Proteomics: The set of proteins expressed by the genetic material of an organism under a given set of environmental conditions.
11 The -omics Revolution in Infertility Metabolomics: The systematic study of the unique chemical fingerprints that specific cellular processes leave behind. Embryomics: The identification, characterization and study of the diverse cell types which arise during embryogenesis.
12 Preimplantation Genetic Diagnosis (PGD) Goal: Identify Genetically Abnormal Embryos IVF/ICSI + Embryo Culture Trophectoderm Biopsy of Blastocyst (~day 5) CCS (comprehensive chromosomal screening) Transfer of Normal Blastocysts/Frozen Embryos Next Step??
13 How Will CRISPR Technology change PGD?? Clustered Regularly Interspaced Short Palindromic Repeats Rapidly remove/add genes using Cas nuclease and synthetic guide RNA Used to modify mosquitos so that they cannot transmit malaria Major potential for embryo gene defect correction Ongoing human research in China, UK, and US on nonviable embryos Huge ethical issues??
14 CRISPR Technology
15 PGD Trophectoderm Embryo Biopsy
16 Comprehensive Chromosomal Screening (CCS) Next Generation Sequencing (NGS) Quantitative PCR (qpcr) Single Nucleotide Polymorphism (SNP array) DNA Fingerprinting Comparative Genomic Hybridization (array CGH) Whole Genome Sequencing (future)
17 Oocyte Cryopreservation Preservation of Oocytes Prior to Fertilization TV Retrieval of Stimulated Oocytes Future: Unstimulated Oocytes with IVM Desiccation and Cryopreservation Delayed Thaw and IVF/ICSI Embryo Culture Transfer of Healthy Embryos
18 Oocyte Desiccation for Cryopreservation
19 Fertility Preservation
20 Fertility Preservation
21 IVF Treatment of Genetic Mitochondrial Disease Mitochondrial DNA Replacement Therapy Performed by Donor Egg Nuclear Transfer 1:200 incidence of pathogenic mtdna mutation Recently approved by British Parliament Currently in active IRB-approved study at OHSU
22 Stem Cell Gamete Production A. Goal is cost-effective, ethically-acceptable source of sperm and oocytes - Reduction of risks with donor gametes - Alternative to somatic cell cloning B. Potential treatment for cancer patients, age-related infertility, and severe male factor C. Reduction of multiple gestation by SET
23 Mouse Embryonic Stem Cell Spermatid NPR Dec. 2004
24 Mouse Blastocysts Fertilized by Embryonic Stem Cell Spermatids NPR Dec. 2004
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26 In Vitro Fertilization (IVF) SART Data: 68,782 IVF babies born in 2014 in U.S. IVF babies now constitute almost 2% of U.S. births Estimated 500,000 IVF babies born in 2015 in world IVF births now almost 4% of births in Europe Estimated >7,000,000 IVF births by Jan Who Knew?????
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28 Polycystic Ovary Disease: A Common Endocrine Disorder in Women Paul Kaplan, M.D. Clinical Professor of Reproductive Endocrinology - OHSU Courtesy Senior Research Associate, Human Physiology University of Oregon
29 Case Presentation Jenn A. 23 Y. O. G0 P0 menarche age 13 BMI 29. Hx of weight problems. Menses Q days. Start BCPS age 15. Moderate acne & hirsutism age Family Hx T2 diabetes and infertility
30 Polycystic Ovary Syndrome A Common Female Endocrine Dysfunction Affects 1 of every ~12 women in U.S. Key Features: Oligo/Amenorrhea Abnormal Androgen Production & Metabolism Probable Genetic Etiology Conveys evolutionary metabolic efficiency? autosomal dominant/variable penetration
31 PCOS - History 1935 Stein-Leventhal Syndrome Observed association of amenorrhea and polycystic ovaries (at surgery) Currently 30,000 published articles on PCOS Now recognized as the leading cause of infertility
32 Irving Stein, M.D. American Journal of Obstetrics & Gynecology , 247.e1-247.e11DOI: ( /j.ajog ) Copyright 2016 Elsevier Inc. Terms and Conditions
33 Michael Leventhal, M.D. American Journal of Obstetrics & Gynecology , 247.e1-247.e11DOI: ( /j.ajog ) Copyright 2016 Elsevier Inc. Terms and Conditions
34 PCOS: A NEW PARADIGM PCOS is a metabolic disorder affecting multiple body systems that requires comprehensive and long-term evaluation and management. John Nestler, M.D. Fertility & Sterility November, 1998
35 PCOS: Evolutionary Benefits Metabolic Thriftiness Maximal caloric conservation Longevity in animal studies Stress-induced ovulation ( LH P/F ) Rate of oocyte atresia ( Insulin levels)
36 How Do Women with PCOS Present? Irregular Menstual Periods Hirsutism Facial Acne Overweight Infertility Acanthosis Nigricans (café au lait spots)
37 Acanthosis Nigricans
38 PCOS: Diagnosis N.I.H. Definition (2 of 2) Oligo/Anovulation Cycles > 35 days apart or < 7 per year Abnormal Androgen Production & Metabolism Clinical (Hirsutism/Acne) or Lab (T, A, DHEA-S) ESHRE/ASRM Rotterdam 2003 (2 of 3) Oligo/Anovulation Androgen Excess Polycystic Ovaries (12 or > follicles/ovary on U/S) Rotterdam ESHRE/ASRM RM Group. Hum Reprod 2004;19: Chang RJ. Am J Obstet Gynecol 2004;191:
39 HYPERANDROGENIC
40 PCOS: CVD Classification Classic (75%) RD/NIH Criteria + Overweight (BMI > 25) 40% Risk of IGT or T2DM by age 40 (5X controls) Dyslipidemia in 70% of Classic PCOS (IR effect) Ovulatory (Lean) (12.5%) Medium risk profile Nonhyperandrogenic (12.5%) Lowest Risk Assessment of Cardiovascular Risk in PCOS. JSEM May2010;95:5.
41 PCOS: Clinical Consequences Endometrial Cancer (3x risk, up to 1/5) Spontaneous Abortion (? LH Effect) Gestational and Type 2 Diabetes (5-7x) Cardiovascular Disease ( LDL HDL) Hypertension Breast Cancer (3-4x risk in limited data) Ovarian Cancer
42 Evaluation of PCOS BMI, Waist Circumference, BP Baseline FSH, LH, TSH, Prolactin Testosterone, DHEA-S 17-OH Progesterone (Follicular a.m.) Fasting Glucose + Insulin/GTT Fasting Lipids & Chemistry Panel Transvaginal Ultrasound of Ovaries
43 String of Pearls in PCOS Transvaginal Ultrasound of the Ovaries
44 PCOS: Insulin Resistance Demonstrated in 60-80% of PCOS 95% in Obese PCOS (BMI > 30) Metabolic Effects: Decreased Hepatic SHBG Production ( Free T) Increased Ovarian Thecal Androgen Production Increased Triglycerides and Adverse Lipid Profile Obesity/Metabolic Syndrome Hypertension High Risk of T2 DM (25%)
45 Metabolic S yndrome: Diagnosis Three or more of the following: Hypertension (130/85 or higher or on meds) Elevated Triglycerides (>150 mg/dl or on meds) Reduced HDL (Less than 50 mg/dl for women) Waist circumference > 35 inches for women Fasting Glucose >100 mg/dl or on meds
46 Metabolic S yndrome: What We K now Occurs in 1/6 (16%) of the general population and 60% of obese men and women. 10% of people with NGT, 40% with IGT, 85% with T2 DM. Prevalence 24% higher in women (40% by age 60) and increases with age. Conveys a high risk of T2 DM and cardiovascular disease. Significant increased risk with PCOS
47 Metabolic S yndrome: Treatment Lifestyle: Diet, Exercise, Weight Loss Correction of Problems: HTN, DM, Lipids Regular monitoring/follow-up? Low-dose aspirin
48 PCOS: Cardiovascular Disease Dyslipidemia Hypertension Impaired Glucose Tolerance/Type2DM Metabolic Syndrome Frequent Positive FH CVD before age 55 Carotid-IMT (10-15% over controls) Carotid Artery Calcification Multivessel CVD (32% vs. 25%) Assessment of Cardiovascular Risk in PCOS. JSEM May2010;95:5.
49 PCOS: Treatment Options Anovulation: Cyclic Progestins, BCPs Prevent D.U.B., Endometrial CA Acne/Hirsutism: BCPs, Spironolactone Contraception: Low-Androgenic BCPS Fertility: Clomiphene, Aromatase Inhibitors, FSH/hMG Weight Loss: Low Calorie ADA Diet Role of Insulin-Sensitizing Medications
50 PCOS: Positive Effects of Insulin Sensitizing Agents SHBG ( Androgen Binding) Testosterone and Androstenedione Triglycerides and LDL Regulation of Menstrual Cycles (30%) Weight Loss (Slow) Increased Sensitivity to Ovulation Meds? Decreased Risk of Miscarriage
51 PCOS: Who to Treat with Metformin in 2015 Obese (BMI>30) PCOS patients Adults Adolescents (who can be compliant) Insulin resistant patients Fasting Insulin > or G/I ratio < 4.5 Young patients (age <30) desiring fertility Patients with impaired glucose tolerance (IGT) or Type 2 D.M.
52 PCOS : Management S ummary Tailor treatment to life stage Induce regular menses Identify & treat endocrinopathies Identify & treat insulin resistance Ongoing regular medical follow-up
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