PREIMPLANTATION GENETIC SCREENING REVIEW. Reproductive Health Science Ltd Dr Michelle Fraser, CEO and MD March 2016

Transcription

1 PREIMPLANTATION GENETIC SCREENING REVIEW Reproductive Health Science Ltd Dr Michelle Fraser, CEO and MD March 2016

2 Forward Looking Statements Any forward looking statements in this presentation have been prepared on the basis of a number of assumptions which may prove incorrect and the current intentions, plans, expectations and beliefs about future events are subject to risk, uncertainties and other factors, many of which are outside Reproductive Health Science Limited s control. Important factors that could cause actual results to differ materially from any assumptions or expectations expressed or implied in this presentation include known and unknown risks. As actual results may differ materially to any assumptions made in this presentation, you are urged to view any forward looking statements in this presentation with caution. This presentation should not be relied on as a recommendation or forecast by Reproductive Health Science Limited, and should not be construed as either an offer to sell or a solicitation of an offer to buy or sell shares in any jurisdiction. 2

3 Number of cycles ART success rate and maternal age 16,000 14,000 12,000 10,000 8,000 6,000 4,000 All initiated cycles Embryo transfer cycles Clinical pregnancies Live deliveries 2,000 0 < >=45 Maternal age Assisted Reproductive Technology (ART) success rates decrease with advancing maternal age Assisted Reproductive Technology in Australia and New Zealand 2013, published September

4 Aneuploidy Typical human cells contain 46 chromosomes arranged in 22 pairs of autosomes numbered 1 to 22 and either XX or XY. An aneuploid cell contains an abnormal number of chromosomes. 4

5 Rate Aneuploidy and maternal age 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0% < >42 Maternal age D3 aneuploidy D5/6 aneuploidy Harton et al 2013 Even younger IVF patients have significant numbers of aneuploid embryos 5

6 Number of transfers Aneuploidy significantly effects outcome Live birth or ongoing pregnancy Unsuccessful 10 0 Euploid embryos 4 Aneuploid embryos 96% of the aneuploid embryos (ie embryos with the incorrect number of chromosomes) failed to implant resulting in an unsuccessful IVF transfer Scott et al

7 Preimplantation genetic screening (PGS) By identifying aneuploid embryos prior to IVF transfer, the chances of an unsuccessful transfer are significantly reduced Embryos can be biopsied at different stages of development, the 8 cell stage (blastomere) usually on day 3 of development through to the blastocyst on day 5/6 and the chromosomal content tested This is known as Pre-implantation Genetic Screening ( PGS ) PGS is a selection tool to assist in the identification of the embryo with the greatest likelihood of viability allowing the prioritisation of transfer 7

8 The clinical intent of PGS To increase IVF implantation rates and ultimately take home baby rates To reduce the number of embryo transfers required to achieve a successful outcome Cost saving Time saving Creates a more positive patient experience than multiple failed cycles The IVF industry is increasingly incorporating PGS in patient management and business planning based on published data and their own experience 8

9 PGS and the age effect With PGS, implantation rates do not decrease until maternal age is >40 years 100% 90% 80% 70% 60% 50% 40% 30% D3 implantation rate D5/6 implantation rate 20% 10% 0% < >42 Maternal age Harton et al

10 PGS in repeat implantation failure patients Clinical outcomes following blastomere biopsy and PGS were improved. 100% 90% 80% 70% 60% 50% 40% PGS No-PGS 30% 20% 10% 0% Clinical pregnancy Implantation Miscarriage Lukaszuk et al

11 No one would knowingly transfer an aneuploid embryo Handyside

12 Array CGH First use of acgh for PGS Hu et al 2004 Fishel et al year old woman, 13 previous failed cycles Polar body biopsy of 7 fertilised oocytes, 2 euploid Day 3 morphology 6 cell grade 3 4 cell grade 2 Both transferred 12

13 24sure Fishel et al

14 X Y Negative Mean of normalized ratios acgh now Illumina 24sure support.illumina.com RHS EmbryoCellect TM rhsc.com.au Chromosome Agilent GenetiSure agilent.com 14

15 Reproductive Health Science (RHS) RHS is a developer of novel products for the analysis of the genetic content of single cells Our lead product EmbryoCellect TM contains: DNA amplification and fluorescent labelling components and A microarray for counting the number of chromosomes in a single human cell This product is being applied to improve the success rate of In-Vitro Fertilisation (IVF) through Pre-Implantation Genetic Screening (PGS) 15

16 The RHS EmbryoCellect TM Kit The end user product comprises the PCR and Labelling Kit and the microarray Manufacturing scale-up of the RHS microarray is uncomplicated and inexpensive The kits are research use only products 16

17 X (Inv) Yp Yq11 Negative Mean of normalized ratios EmbryoCellect TM - improved, simpler PGS Male Equal number of chromosomes Outlier; missing a copy of chromosome 15 Chromosome RHS microarray results generated from a biopsy of an aneuploid embryo (45,XY,-15) By capturing the results from an entire chromosome in a single microarray feature, the aneuploidy result is simply obtained. Having eight replicates for each chromosome allows an assessment of the quality of the array result and overcomes interference from debris, bubbles or other hybridisation artefacts. 17

18 EmbryoCellect TM validation RHS has validated the performance of EmbryoCellect TM using single and small numbers of cells from known euploid and aneuploid cell lines biopsies from embryos previously tested with other PGS kits Analysing the same amplified samples using the EmbryoCellect TM microarray and Next Generation Sequencing, and through successful clinical outcomes for patients with previous multiple IVF cycle failures 18

19 EmbryoCellect TM capabilities Quality, robust whole genome amplification that provides a solid foundation for genetic analysis Each embryo treated as precious "with flexibility of integrated clinical and scientific team Straightforward chromosomal aneuploidy report without the complication of interpreting unnecessary additional genetic information Next Generation Sequencing offers no additional information over microarrays for aneuploidy detection, the PCR being the limiting step and amplified single cell DNA effectively dampened down 29

20 EmbryoCellect TM Competitive Advantages Robust single cell validation conducted by RHS prior to clinical use to ensure product accuracy Embryo testing externally validated against competing PGS products providing end user feedback and product enhancements and evidence of equivalent performance Ease of use allowing clinics new to PGS to introduce the system with minimal upskilling Simple interpretation not reliant on complicated algorithms providing faster, clearer results No complex genetic counselling limited ethical and legal risks accurate aneuploidy information without the complication of higher than needed resolution Cost competitive pricing allowing broader patient access 20

21 Next Generation Sequencing (NGS) First babies born in the USA in the Summer of IVF cycles 5 blastocysts in cycle 1 2 blastocysts in cycle 2 eset Both resulted in live births 31

22 Next Generation Sequencing PGS uses low pass sequencing, which gathers a small amount of sequence information across the genome Limited coverage breadth and depth 0.04x Analysing multiple embryos together increases throughput and keeps cost lower The detection of genetic errors of clinical significance and SNPs requires read depths of at least 30x for accuracy It is not possible to accurately combine PGS and PGD unless using a higher resolution sequencing approach, which increases cost and raises the risk of detecting incidental findings or variants of unknown clinical significance, presenting ethical and medicolegal concerns The biggest limitation remains in the PCR amplification step, where the sample needs to be accurately and comprehensively copied Combined PGS (low pass) and PGD (deep) sequencing is not possible with current protocols 33

23 Microarrays or Next Generation Sequencing? What are the drivers? Increased throughput suited to larger IVF clinics or centralised service providers Possible reduced hands on time Cost savings when analysing 24 samples in a batch The promise of future improved resolution compared to microarrays, currently there is no significant difference Microarrays are the better option for clinics wanting simpler results and/or that don t have the volumes for NGS or when a fresh embryo transfer is required Lower set-up costs Smaller batch sizes so no need for banking With current technologies, the 6 picograms of DNA that is in a single cell is not enough to achieve a whole genome sequence 36

24 RHS and Next Generation Sequencing X MiSeq reads for 48,XY,+2,+21 single cells normalized to 46,XX single cells RHS have validated that the single cell aneuploidy detectable using EmbryoCellect TM is also detectable using sequencing RHS plans to launch a lysis and whole genome amplification kit for this purpose This kit would also have application in single cell genetic analysis beyond the IVF market 35

25 Equipment needed Molecular biology equipment eg PCR machine, gel imaging system, laminar flow, pipettes Standard laboratory equipment eg freezer, fridge Microarrays NGS Specialised equipment Microarray scanner, water bath, slide warmer Sequencer, library preparation equipment, DNA quantification equipment 38

26 Reprogenetics; an example of change 100% 90% 80% 70% 60% 50% 40% 30% 20% NGS day 5 NGS day 3 acgh day 5 acgh day 3 acgh FISH 12 FISH 5-9 Vitrified/frozen 10% 0% Reproduced with permission from Dagan Wells 39

27 Expertise needed World standard IVF laboratory Extended embryo culture to allow blastocyst culture Biopsy Tube transfer to ensure test success Molecular biology experience will assist in rapid validation and integration Clinical genetics experience to understand and accurately interpret results Patient information preparation and counselling for informed consent 40

28 Whole Genome Amplification The most important determinant of accuracy for genetic analysis of single or small numbers of cells is the whole genome amplification. This is regardless of whether NGS or microarrays are used for the analysis There are 4 fundamental approaches; Linker Adapter-PCR eg SurePlex DOP-PCR eg RHS MDA eg RepliG, SureMDA MALBAC eg Yikon Genomics The upcoming launch of RHS s lysis and whole genome amplification kit will be the first DOP-PCR kit on the market 34

29 The global IVF market Country IVF cycles per year (% of global) Population (millions) IVF cycles per million people RHS distributor Russia 80,000 (4.3%) BioChem Mack Iran 35,000 (1.9%) Vitco UAE and Saudi Arabia 8,000 (0.4%) Reprolab Turkey 45,000 (2.4%) Tani Medikal India 80,000 (4.3%) 1, SAR Healthline Indonesia 3,500 (0.2%) Direct South Africa 5,500 (0.3%) Delfran Israel 32,000 (1.7%) 8 3,902 Al-Rad Australia 62,000 (3.3%) 23 2,605 Direct - service China 250,000 (13.3%) 1, Miaoquan USA 163,000 (8.7%) The rest of the World, predominantly Europe, make up the remaining 40.8% of the global IVF market Data sourced from publications and RHS distributor supplied information 41

30 The EmbryoCellect TM distributors The distributor relationships provide RHS with access to their customer networks that have been built over many years EmbryoCellect TM is part of a catalogue of IVF products with established channels Some RHS distributors also supply the necessary equipment and have in-house technical expertise to assist with new laboratory set-up and local customer support 30

31 Sales strategy in-field evaluations Evaluation by select key clinics RHS supplies small numbers of EmbryoCellect TM kits to select key account IVF clinics to obtain end user feedback Local marketing considerations individually addressed by working closely with the clinics and distributors to provide RHS with an indication of the required distributor and clinical users in-market support Short term studies, predominantly benchmarking against other PGS platforms, by clinics are underway 43

32 Sales strategy training courses RHS held its second EmbryoCellect TM training course at our facilities in November 2015 Attended by representatives from 4 clinics from China and one from Australia Provided attendees with hands-on training, laboratory set-up advice and access to RHS expertise The first training course for 2016, will be held in March, with attendees from Japan and Brazil 44

33 RHS PGS service business Established to meet customer needs Requires shipment of samples to RHS and availability at the IVF clinic of an embryologist capable of undertaking biopsy and vitrification Utilises RHS expertise and infrastructure to support clinics who are establishing PGS Enhances EmbryoCellect TM uptake and awareness Represents an additional revenue stream to RHS Current service agreements with two Australian and one overseas clinic 45

34 EmbryoCellect TM global marketing 2016 International conferences to attend Asia Pacific Initiative on Reproduction (ASPIRE) Jakarta, April 2016, exhibitor American Society for Reproduction Medicine (ASRM) Salt Lake City, October 2016 exhibitor European Society Human Reproduction and Embryology (ESHRE) Helsinki, July 2016, exhibitor Pre-implantation Genetic Diagnosis International Symposium (PGDIS) Bologna, May 2016, exhibitor 2016 National conferences to attend Fertility Society of Australia (FSA) Perth September 2016 Scientists in Reproductive Technologies (SIRT) Adelaide, April/May

35 The Opportunity : PGS Market Forecasts 2013 data Forecast 2019 Global IVF cycles per annum 1.7m* 3m (annual growth 10%) Global PGS cycles per annum 51, ,000 (3% of IVF market) ** (20% of IVF market)*** Number of tests per IVF cycle (average, estimate) Number of PGS tests per annum , m *Estimated based on there being a reported 1.5m IVF cycles in 2010 and 10% growth per annum ** Illumina estimate Jan 2014 *** RHS estimate 35

36 The increasing use of PGS RHS market intelligence suggests that 20% of US and 30% of Australian IVF cycles use PGS Virtus increased their number of PGS cycles by 54% in the last half year 36

37 Patient decision-making A US patient questionnaire identified 3 significant factors influencing their decision to undertake a PGS cycle The additional cost of screening The provider information and influence Social support or acceptance from partner/family/friends By providing an accurate, cost effective, easy to use and interpret PGS solution, RHS expects EmbryoCellect TM to become a significant part of the expanding global PGS market Gebhart et al 2016 Reference 12 51

38 Profit & Loss year to Dec 2015 First EmbryoCellect TM and Services sales were recognised in 2015 $73k Additional deferred sales $27k Other income of $360K (R&D, EMDG & interest) The Company contains operating costs whilst commercialising EmbryoCellect TM Average monthly cash burn remains = $160,000 38

39 Financial Position Cash at bank 31 December m EMDG grant first instalment expected March 0.04m = Available funds 1.50m + EMDG 2nd instalment expected mid R&D Tax Refund being prepared Monthly operating cash requirements remain < 0.16m Available cash is sufficient to fund 9 months, before taking into account further sales and receipt of the 2015 R&D Tax Refund Additional client orders have been received in Feb/March Orders are expected to increase following client evaluations 39

40 The RHS goal RHS estimates that 20% of global IVF market will be using PGS by The current uptake varies between countries with the Australian PGS market being one of the highest, currently estimated at 30% of IVF cycles RHS aiming for 20% of overall global PGS market by Our current product reach is in over 30% of the global IVF market. 40

41 Appendix of information slides RHS background and corporate overview The RHS products Intellectual Property Key Competitors IVF and PGS support information Comparative workflows for microarrays and Next Generation Sequencing 41

42 Background of RHS Commenced single cell genomics R&D in 2004 Initial venture capital investment Oct 2007 $4.4m invested and $1.3m grant funding leveraged Listed on ASX at $0.20 in April 2014 Located within the BioSA Incubator at Thebarton, South Australia Dedicated laboratory and office space Manufacturing capability with scope for scale-up Established network of commercial partners 42

43 Corporate Overview Exclusively in-licensed patent family Primary value driver Single cell genetic analysis know-how Primary value driver Key assets Granted patent family in most key territories to method for detecting chromosomal abnormalities Clinical impact in the IVF market Kits developed for the PCR amplification of multiple and single cells for microarrays and sequencing Global product sales and key partnerships ASX Code 31 December 2015 data Shares on Issue Share Price (on small trades) RHS 59m Options 7.3m Market Capitalisation Cash at 31 Dec c $4.7m $1.46m Recent Corporate Highlights Service provision commenced with 2 clinics (and a 3 rd service agreement late 2015) Distributor agreements with 8 companies covering over 30% of the global IVF market RHS continues EmbryoCellect TM training 43

44 Board of Directors Dr David Brookes Non-Executive Chairman Dr Michelle Fraser Managing Director and CEO Ms Sue MacLeman Non-Executive Director Mr Fabian Dwyer Non-Executive Director Mr Johnathon Matthews Non-Executive Director Emeritus Professor Colin Matthews AO Alternate Non-Executive Director Director of Atcor Medical Holdings Ltd (ACG:ASX); medical practitioner & biotechnology consultant; MBBS; FACRRM; FAICD PhD (molecular biology); Grad Dip Science & Technology Commercialisation; GAICD Distinguished career in biotechnology with wealth of industry experience; has been CEO and a Board member of a number of publicly-listed companies in both the USA and Australia; BPharm; MMktg; MLaw; FACPP; FAICD Experienced senior sales and marketing executive with several pharmaceutical companies and has held CEO roles in research and commercialisation companies; BPharm; MBA; MRPharmS; FAICD Former executive director of The Pipette Company; previously held positions at Australian Treasury, ASX and Commonwealth Bank; BEc; BComm; LLB; GAICD Inaugural director of RHS, Single Cell Pty Ltd, Flinders IVF; cofounder and former director of The Pipette Company; former Director ReproMed; former Chairman of Research Committee and Board member of Channel 7 Research Foundation. Chairman RHS Clinical &Scientific Advisory Committee 44

45 The RHS products What are they? How do they work? What is their potential? Lead product EmbryoCellectTM for IVF PGS (Pre-implantation Genetic Screening) 57

46 Testing Single Cells Start with a single cell RHS PCR System Polymerase Chain Reaction (PCR) Generate lots of copies of the cell s DNA using PCR to create enough test material RHS Microarray Interrogate the cell to gain information on the entire entity Count the number of chromosomes in the cell 46

47 RHS Products Lead product EmbryoCellect TM IVF kit RHS microarray RHS PCR kits Cell lysis and DNA amplification Fluorescent labelling Additional product Next Generation Sequencing 47 59

48 Intellectual Property Microarray patent family; RHS has exclusively in-licensed a patent family from The University of Adelaide Patent has been granted/allowed in the United States of America, Australia, New Zealand, Canada, Europe and China and is imminent in Hong Kong RHS has more than 10 years of know-how, providing significant expertise in the genetic analysis of low copy number DNA and single cells These technologies have broader applications 48

49 Key Competitors in Single Cell Analysis Technology DNA amplification Microarrays Sequencing Company Rubicon Genomics Inc, Sigma-Aldrich Co LLC, Qiagen NV, Yikon Genomics Illumina Inc*, Agilent Technologies Inc, Natera Inc, Oxford Gene Technologies Life Technologies Corp**, Illumina Inc, Fluidigm Corp *In September 2012, BlueGnome was acquired by Illumina for USD $95.5m including USD $88m in initial cash payments. At that time, Bluegnome were generating USD$17m in revenues and $1m in profits **In February 2014, Thermo Fisher Scientific acquired Life Technologies RHS products compete across the DNA amplification, microarray and sequencing markets 49

50 IVF A clear need for improvement 80,000 70,000 71,516 Australian and New Zealand ART ,000 50,000 40,000 30,000 54,926 70% of embryo transfers do not result in clinical pregnancy 20,000 10,000 17,054 12,997 0 Cycles initiated Embryo transfers Clinical pregnancies Live births Assisted Reproductive Technology in Australia and New Zealand 2013, published September

51 US IVF success rates 250,000 United States of America ART , , , , ,243 51% of embryo transfers do not result in clinical pregnancy 50,000 65,652 53,286 0 Cycles initiated Embryo transfers Clinical pregnancies Live births CDC National Summary 2013, published November

52 UK IVF success rates 70,000 United Kingdom ART ,000 50,000 40,000 57,522 39,874 65% of embryo transfers do not result in clinical pregnancy 30,000 20,000 10,000 13,788 11,963 0 Cycles initiated Embryo transfers Clinical pregnancies Live births HFEA Fertility Treatment in 2013: trends and figures, published December

53 Single embryo transfer vs implantation 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0% Australia USA UK eset Non-eSET Implantation rate per transfer There are 2/3 more multiple embryo transfers in the US compared to Australia but there is only 1/3 increase in implantation rate Assisted Reproductive Technology in Australia and New Zealand 2013, published September 2015 CDC National Summary 2013, published November 2015 HFEA Fertility Treatment in 2013: trends and figures, published December

54 US rate of single embryo transfer 100% 90% 80% 70% 60% Transferring more than one embryo is common in the US. Although it increases implantation rates, it is associated with multiple births and significant clinical implications 50% 40% 30% 20% 10% 31% 24% 18% 14% 10% 7% 0% < >44 Maternal age CDC National Summary 2013, published November

55 IVF embryo aneuploidy rate is significant Clinical study of 146 couples Mean maternal age 34.0 ±4.4 years 232 IVF embryos transferred blindly prior to PGS results being generated Euploid Aneuploid More than 40% of the embryos were aneuploid Scott et al

56 Number of embryos D3 vs D5 PGS results The aneuploidy rate in blastocysts is lower than in blastomeres probable selection during extended culture Euploid Aneuploid 10 0 Blastomere Blastocyst Scott et al

57 Embryo transfers D3 vs D5 transfer results Implantation and live birth rates are higher with euploid embryos after blastocyst biopsy than after blastomere biopsy Live birth or ongoing pregnancy 10 0 Euploid blastomere 1 Aneuploid blastomere Euploid blastocyst 3 Aneuploid blastocyst Scott et al

58 Donor oocyte outcomes 39% of blastocysts derived from donor oocytes were aneuploid. Clinical outcomes improved with PGS, particularly the miscarriage rate 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0% Clinical pregnancy Ongoing pregnancy Implantation Miscarriage PGS Control Haddad et al

59 The influence of frozen transfers 100% Clinical outcomes were comparable between fresh and frozen embryo transfers without PGS but improved following PGS and frozen embryo transfer. It is the use of PGS that improved IVF outcomes. 90% 80% 70% 60% 50% 40% 30% No-PGS fresh No-PGS FET PGS-FET 20% 10% 0% Implantation Live births per transfer Live births per implantation Lee et al

60 Microarray Workflow Sample collection Time Module 1a: RHS WGA (1st round DOP-PCR) Lysis 30 min set-up 15 min Whole Genome Amplification (WGA) Labelling PCR 2 hr 30min 30 min 15 min 1 hr 30 min - 1 hr Hybridisation 3 hr to overnight Washing and Scanning min 1 hr 27

61 NGS Workflow Sample collection Module 1a: RHS WGA (1st round DOP-PCR) Lysis Time 30 min set-up 15 min Whole Genome Amplification (WGA) 2 hr 30min 30 min QC DNA quantification and dilution 20min Library preparation 65min DNA Clean-up 30min Library normalization and pooling 1 hr 30min Sequencing run ~ 5hr (1x36bp) VeriSeq is marketed as 12hr protocol Read alignment and analysis 1 hr 32

62 AT THE FOREFRONT OF SINGLE CELL GENETIC ANALYSIS Reproductive Health Science Ltd Dr Michelle Fraser, CEO and MD