Autologous mitochondrial injection to increase
energy in oocytes
Robert Casper, M.D., F.R.C.S.C.
University of Toronto
TCART Fertility Partners
Global Demand for Fertility Treatments is Growing
2
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Approximately 1.5 million in vitro fertilization (IVF) cycles
reported per year worldwide
•
IVF has been in clinical practice for 35+ years
•
Despite significant gains in the live birth rate in good
prognosis patients, there are few treatment options for
repeated IVF failures
Preclinical Support for Safety and Efficacy of
Mitochondrial Transfer
3
New Option:
AUGMENTSM (Autologous Mitochondrial Treatment)
Convincing evidence that adequate oocyte mitochondrial
activity is necessary for embryo development to blastocyst
and for successful implantation1
Goals:
• Improve oocyte and embryo quality
• Increase healthy live birth rate
1.
4
Chappel, Obst and Gyn Intl 2013; Tilly et al, Cell Metab 2013
Discovery of Egg Precursor Cells
1. White, Y. A. R. et al. Oocyte formation by mitotically active germ cells purified from ovaries of reproductive-age women.
Nat. Med. 18, 413–421 (2012).
2. Woods, D. C. & Tilly, J. L. Isolation, characterization and propagation of mitotically active germ cells from adult mouse
and human ovaries. Nat. Protoc. 8, 966–988 (2013).
3. Zou, K. et al. Production of offspring from a germline stem cell line derived from neonatal ovaries. Nat. Cell Biol. 11,
631–636 (2009).
4. Zhang, Y. et al. Production of transgenic mice by random recombination of targeted genes in female germline stem
cells. J. Mol. Cell Biol. 3, 132–141 (2011).
5. Zhou, L. et al. Production of fat-1 transgenic rats using a post-natal female germline stem cell line. Mol. Hum. Reprod.
20, 271–281 (2014).
6. Wolff, E. F. et al. Oogonial stem cells generate mature oocytes in an autologous Rhesus macaque transplantation
model. Reprod. Sci. 21, 119A (2014).
7. Anderson, R. A. et al. Evaluation of oogonial stem cells and neo-oogenesis in ovaries of girls and women with Turner
syndrome: a pilot study. Hum. Reprod. 2013; 28 (Suppl.): 52 (2013).
8. Dunlop, C. E., Bayne, R. A., McLaughlin M., Telfer, E. E. & Anderson, R. A. Isolation, purification, and culture of oogonial
stem cells from adult human and bovine ovarian cortex. Lancet 383, S45 (2014).
9. Zou, K., Hou, L., Sun, K., Xie, W. & Wu, J. Improved efficiency of female germline stem cell purification using Fragilisbased magnetic bead sorting. Stem Cells Dev. 20, 2197–2204 (2011).
10. Park, E. S., Woods, D. C., White, Y. A. R. & Tilly, J. L. Oogonial stem cells isolated from ovaries of adult transgenic
female mice generate functional eggs and offspring following intraovarian transplantation. Reprod. Sci. 20, 75A–76A (2013).
11. Imudia, A. N. et al. Comparative gene expression profiling of adult mouse ovary-derived oogonial stem cells supports a
distinct cellular identity. Fertil. Steril. 100, 1451–1458 (2013).
12. Park, E. S., Woods, D. C. & Tilly. J. L. Bone morphogenetic protein 4 (BMP4) promotes mammalian oogonial stem cell
differentiation via SMAD1/5/8 signaling. Fertil. Steril. 100, 1468–1475 (2013).
13. Xie, W., Wang, H. & Wu, J. Similar morphological and molecular signatures shared by female and male germline stem
cells. Sci. Reports 4, 5580 (2014).
14. Khosravi-Farsani, S., Amidi, F., Roudkenar, M. H. & Sobhani, A. Isolation and enrichment of mouse female germ line
stem cells. Cell J. 16, 406–415 (2015).
15. Park, E. S. & Tilly, J. L. Use of DEAD-box polypeptide 4 (Ddx4) gene promoter-driven fluorescent reporter mice to
identify mitotically active germ cells in postnatal mouse ovaries. Mol. Hum. Reprod. 21, 58–65 (2015).
5
Egg Precursor Cell Mitochondria are Similar to
Oocyte Mitochondria
• Egg precursor cells are unipotent, germline cells in
the outer cortex of the ovary
Human Brain Cell
Human Oocyte
Human Egg PC
Human Egg Precursor Cell
500 nm
Human brain: http://dx.doi.org/10.1093/brain/awu138, 2329-2345 2014
Human oocyte: Nottola et al., Hum Reprod 22:1123–1133, 2007
Human egg precursor cell: Harvard Medical Schoool
6
500 nm
500 nm
Autologous Mitochondrial Transfer Possible
Nucleus
Ovarian Cortex
Egg Precursor Cells
1
• Maintains mitochondrial homoplasmy
and nuclear and mitochondrial DNA
communications
1.
7
White et al., 2012
Mitochondria
(maternally inherited)
AUGMENTSM Process
Ovarian Tissue
Biopsy
1
1.
8
Controlled Ovarian
Hyperstimulation
ICSI
Oocyte Retrieval
Embryo Transfer
Mitochondria
1
Oocyte precursor
cells isolated
Performed by OvaScience
Oocyte precursor
cell mitochondria
isolated
Many Conditions are Prognostic for Live Birth
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•
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Age
Oocyte/embryo quality
Antral follicle count
Hormone levels: FSH (follicle stimulating hormone), AMH (antimullerian hormone)
BMI (body mass index)
Environmental factors
Previous diagnoses: diminished ovarian reserve, endometriosis, etc.
Previous failed IVF
Lack of agreement on how to rank significance of risk factors
9
Baseline Clinical Pregnancy Rate
10
Metric
Previous IVF History Data
Number of patients
34
Average current age
36.0 (Range: 26-44)
Background/Diagnoses
Poor oocyte & embryo quality with one of
the following diagnoses:
Diminished Ovarian Reserve, Ovulatory
Dysfunction, Polycystic Ovarian Syndrome,
Tubal Factor, Endometriosis, Unexplained
Total previous IVF cycles initiated
71
Average cycles per patient
2 (Range: 1-5)
Total previous embryo transfers
(fresh & frozen)
79
Historical clinical pregnancy rate:
• per cycle initiated
• per embryo transfer
11%
10%
(8/71)
(8/79)
Historical live birth rate:
• per cycle initiated
• per embryo transfer
1.4%
1.3%
(1/71)
(1/79)
Clinical Experience with AUGMENTSM
Metric
AUGMENT
Number of patients
34
Average current age
36.0 (Range: 26-44)
Background/Diagnoses
Poor oocyte & embryo quality with one of
the following diagnoses:
Diminished Ovarian Reserve, Ovulatory
Dysfunction, Polycystic Ovarian Syndrome,
Tubal Factor, Endometriosis, Unexplained
Total mitochondrial injection cycles initiated
34
Average cycles per patient
1
Total embryo transfers (fresh & frozen)
26
Clinical pregnancy rate:
• per cycle initiated
• per embryo transfer
35%
46%
(12/34)
(12/26)
Ongoing clinical pregnancy and live birth rate:
• per cycle initiated
• per embryo transfer
26%
35%
(9/34) includes 1 live birth
(9/26) includes 1 live birth
9 patients have 23 total frozen embryos remaining for transfer
11
Clinical Experience with AUGMENTSM
Total patients initiated
treatment
34
Patients completed cycles with
mitochondrial injection
30
Patients with embryo transfers
(3 had multiple transfers)
22
3 no fertilization, 4 arrested
embryo development, 1 pending
frozen embryo transfer
Number of embryo transfers
26
(42 total transferred embryos)
Positive pregnancy tests
15
(confirmed by quantitative hCG)
Clinical pregnancies
(confirmed by ultrasound)
Ongoing clinical pregnancies
and live births*
12
1 no sperm obtained, 1 no oocytes
retrieved, 2 oocyte banking
* Includes 1 live birth
3 chemical pregnancies
12
3 spontaneous abortions
9
Clinical Experience Shows Improved Pregnancy
Rates Compared to Baseline
13
SM
Previous IVF
History
AUGMENT
Total cycles initiated
71
34
Average cycles initiated per patient
2
1
Total embryo transfers (fresh & frozen)
79
26
Clinical pregnancies
8
12
Clinical pregnancy rate per cycle initiated
11%
35%
Clinical pregnancy rate per embryo
transfer
10%
46%
Ongoing clinical pregnancies and live
births (rate per cycle initiated)
No ongoing and
1 live birth (1%)
9 including 1 live
birth (26%)
Conclusion
•
Clinical experience with AUGMENTSM during ICSI appears
promising
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–
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Women included in this clinical experience were poor
prognosis for live birth
–
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14
One live birth and 8 ongoing clinical pregnancies
Ongoing clinical pregnancy rate of 26% per cycle initiated and 35%
per embryo transfer
Failed previous IVF cycles, poor embryo development, and 1.4% live
birth rate per previous cycle
Nine patients have frozen embryos remaining for transfer
Acknowledgements
TCART Clinical
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Dr Yaakov Bentov
Dr Paul Chang
Dr Dan Nayot
Deborah Myers
Caroline Lux
First Steps
• Dr Marjorie Dixon
• Dr Fay Weisberg
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TCART Lab
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Dr Navid Esfandiari
Dr Julia Szeptycki
Dennis De la Cruz
Khalid Rao