Does blastocyst mitochondrial DNA content affect miscarriage rate in patients undergoing single euploid frozen embryo transfer?

Abstract Introduction Tissue engineering using human induced pluripotent stem cells-derived cardiomyocytes (hiPSCs-CMs) is one of the potential tools to replicate human heart in vitro. Although there are many publications on 3 dimensional (3D) heart tissues (1), these tissues show fetal like phenotypes. For that reason, several maturation methods such as electrical stimulation and mechanical stress have been investigated (2, 3). However, these methods have been inadequate in differentiating fetal like phenotype tissue from adult tissues. Previously, we identified a novel compound, T112, which induced hiPSCs-CMs maturation from approximately 9,000 compounds using Troponin I1-EmGFP and Troponin I3-mCherry double reporter hiPSCs-CMs. This compound enhanced morphological and metabolic maturation of hiPSCs-CMs via estrogen-rerated receptor gamma activation Purpose We hypothesized that our novel compound, T112, in combination with mechanical stress could result in further maturation of 3D heart tissue. Therefore, our specific aim is to develop a novel maturation method applicable to genetic disease model of HCM using 3D heart tissue combined with T112. Methods We constructed 3D heart tissue mixed with fibroblast and double reporter hiPSCs-CMs by the hydrogel methods using Flex cell system®. We added T112 with or without mechanical stretching to 3D tissue from 7 to 15 days after 3D heart tissue was constructed. Then we measured maturation related phenotype such as sarcomere gene expression, mitochondrial DNA content and cell size. Results Similar to hiPSCs-CM, the addition of T112 to the constructed 3D heart tissue significantly increased TNNI3 mRNA compared to that of DMSO. Furthermore, T112 treated 3D heart tissue showed increased cell size and oblong shape. Next, in order to promote more maturation of 3D heart tissue, we performed mechanical stretching with the addition of T112. The combination of T112 with mechanical stretching showed higher expression of mCherry, a reporter protein for TNNI3 expression, and higher isotropy of sarcomere alignment in 3D heart tissue than that with the static condition. Furthermore, 3D heart tissue in the treatment of T112 with or without mechanical stretching showed higher mitochondrial DNA content compared to the respective DMSO controls. Interestingly, we applied this combination method to hiPSCs carrying MYH7 R719Q mutation which is known to cause hypertrophic cardiomyopathy, and the 3D heart tissue composed of cardiomyocytes derived from mutant iPSCs demonstrated increased sarcomere disarray compared to isogenic wild-type 3D heart tissue. Conclusion These results suggest that the combination of T112 and mechanical stretching promotes metabolic and structural maturation of 3D heart tissue and would be useful for creating a HCM disease model. Funding Acknowledgement Type of funding source: Private company. Main funding source(s): T-CiRA project, Takeda Pharmaceutical Company Limited

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The effects of the day of trophectoderm biopsy and blastocyst grade on the clinical and neonatal outcomes of preimplantation genetic testing–frozen embryo transfer cycles

Zygote ◽

10.1017/s0967199421000435 ◽

2021 ◽

pp. 1-6

Author(s):

Linjun Chen ◽

Zhenyu Diao ◽

Jie Wang ◽

Zhipeng Xu ◽

Ningyuan Zhang ◽

...

Keyword(s):

Birth Weight ◽

Genetic Testing ◽

Embryo Transfer ◽

Live Birth ◽

Frozen Embryo Transfer ◽

Neonatal Outcomes ◽

Miscarriage Rate ◽

Birth Rates ◽

Preimplantation Genetic Testing ◽

Live Birth Rates

Summary This study analyzed the effects of the day of trophectoderm (TE) biopsy and blastocyst grade on clinical and neonatal outcomes. The results showed that the implantation and live birth rates of day 5 (D5) TE biopsy were significantly higher compared with those of D6 TE biopsy. The miscarriage rate of the former was lower than that of the latter, but there was no statistically significant difference. Higher quality blastocysts can achieve better implantation and live birth rates. Among good quality blastocysts, the implantation and live birth rates of D5 and D6 TE biopsy were not significantly different. Among fair quality and poor quality blastocysts, the implantation and live birth rates of D5 TE biopsy were significantly higher compared with those of D6 TE biopsy. Neither blastocyst grade nor the day of TE biopsy significantly affected the miscarriage rate. Neonatal outcomes, including newborn sex, gestational age, preterm birth, birth weight and low birth weight in the D5 and D6 TE biopsies were not significantly different. Both blastocyst grade and the day of TE biopsy must be considered at the same time when performing preimplantation genetic testing–frozen embryo transfer.

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Clinical application of plasma mitochondrial DNA content in lung cancer patients

Annals of Oncology ◽

10.1093/annonc/mdy425.007 ◽

2018 ◽

Vol 29 ◽

pp. ix153

Author(s):

L. Zhang

Keyword(s):

Lung Cancer ◽

Mitochondrial Dna ◽

Cancer Patients ◽

Clinical Application ◽

Dna Content ◽

Lung Cancer Patients ◽

Mitochondrial Dna Content

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Platelet contamination causes large variation as well as overestimation of mitochondrial DNA content of peripheral blood mononuclear cells

Annals of Clinical Biochemistry International Journal of Laboratory Medicine ◽

10.1258/acb.2008.008008 ◽

2008 ◽

Vol 45 (5) ◽

pp. 513-514 ◽

Cited By ~ 35

Author(s):

M. Urata ◽

Y. Koga-Wada ◽

Y. Kayamori ◽

D. Kang

Keyword(s):

Mitochondrial Dna ◽

Peripheral Blood Mononuclear Cells ◽

Dna Content ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Peripheral Blood Mononuclear ◽

Mitochondrial Dna Content ◽

Blood Mononuclear Cells

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Low Tumor Mitochondrial DNA Content Is Associated with Better Outcome in Breast Cancer Patients Receiving Anthracycline-Based Chemotherapy

Clinical Cancer Research ◽

10.1158/1078-0432.ccr-17-0032 ◽

2017 ◽

Vol 23 (16) ◽

pp. 4735-4743 ◽

Cited By ~ 8

Author(s):

Marjolein J.A. Weerts ◽

Antoinette Hollestelle ◽

Anieta M. Sieuwerts ◽

John A. Foekens ◽

Stefan Sleijfer ◽

...

Keyword(s):

Breast Cancer ◽

Mitochondrial Dna ◽

Cancer Patients ◽

Dna Content ◽

Breast Cancer Patients ◽

Mitochondrial Dna Content

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Abstract 381: Prolonged Renal Failure Leads to Reduced Number of Active Cardiac Mitochondria in a Rat Model for Long Term Chronic Kidney Disease

Circulation Research ◽

10.1161/res.117.suppl_1.381 ◽

2015 ◽

Vol 117 (suppl_1) ◽

Author(s):

Einat A Hertzberg-Bigelman ◽

Michal Entin-Meer ◽

Genya Aharon-Hananel ◽

Ann Saada ◽

Ran Levy ◽

...

Keyword(s):

Chronic Kidney Disease ◽

Mitochondrial Dna ◽

Kidney Disease ◽

Cardiac Function ◽

Rat Model ◽

Dna Content ◽

Cardiac Mitochondria ◽

Mitochondrial Content ◽

Mitochondrial Dna Content

Objectives - Cardiorenal syndrome type 4 is characterized by primary chronic kidney disease (CKD) leading to an impairment of cardiac function. We recently showed a reduced expression of several cardiac mitochondrial genes in short-term CKD rat model. We aimed to evaluate whether cardiac mitochondrial structure and function is modified in long-term CKD and if so, to characterize the potential associated mechanisms. Methods - Lewis rats underwent 5/6 nephrectomy for induction of CKD. Upon necroscopy, eight months later, cardiac sections were analyzed by histology and electron microscopy (EM). Mitochondrial DNA content was determined by the mitochondrial gene, cytochrome B. Mitochondrial content was assessed by citrate synthase (CS) activity in tissue homogenate and respiratory chain function was determined by the activity of complexes I-IV in isolated mitochondria. The levels of PGC1a, a transcription factor for mitochondrial biogenesis, Angiotensin II type 1 receptor and cytosolic cytochrome C were assayed by western blot. Cytokine serum profile was determined by microarray. Results - Long-term CKD leads to cardiac hypertrophy and increased interstitial fibrosis. EM analysis revealed a massive spatial disarrangement accompanied by a considerably increased volume of swollen-damaged mitochondria in CKD hearts (32±3%, n=5, 48±6%, n=4; respectively; p<0.05). Total mitochondrial DNA content was decreased in cardiac tissue of CKD rats. Concomitantly, active mitochondrial content was significantly reduced. Conversely, no differences were observed in respiratory chain enzymes’ functions (complexes I-IV) in isolated active mitochondria. Moreover, inflammatory response and activation of Renin-Angiotensin-Aldosterone-System (RAAS) were detected in the CKD setting. Conclusion - CKD results in a marked reduction of active mitochondria in the heart. Inflammatory cytokines and RAAS, may set a deleterious environment to cardiac mitochondria, as suggested in non-CKD models. The data may represent a significant milestone in the personalized medicine strategy for treating CKD patients who present with normal cardiac function accompanied by positive biomarkers for cardiac mitochondria damage.

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