There are a variety of similarities, as well as some very important differences, for the prenatal molecular diagnostic landscape in Europe compared to the situation in the U.S. or other parts of the world. Arrays and next-gen sequencing are playing an
increased role, but these are not the only technologies being applied to this field. There has been a dramatic decline in the level of invasive testing, predominantly because of the widespread introduction of non-invasive screening based on cell-free
DNA. In Europe, where samples are more likely to be evaluated locally, there is a wider range of options available for cell-free DNA testing compared to other parts of the world, where centralized testing is more prevalent.
While cell-free DNA testing moves beyond just aneuploidy testing; including more testing for sub-chromosomal genetic conditions, these changes may involve higher costs as well as additional questions for false positives and issues for implementation.
At the same time, those researchers working to commercialize an alternative to cell-free DNA, specifically the non-invasive isolation and analysis of fetal cells from maternal blood, are moving into clinical trials prior to being able to commercialize
this approach. It seems clear that before too long these two techniques will both be on the market and will compete with each other. A range of issues related to implementation of newer testing in the prenatal setting will be covered.
Final Agenda
MONDAY, 10 APRIL
08:00 Short Course Registration and Morning Coffee
12:00 Main Conference Registration
13:00 Chairperson’s Opening Remarks
13:10 Chromosomal Microarray in Prenatal Diagnostics – Lessons from More Than 1500 Analyses
Ida Vogel, M.D., Head of Clinical Genetics, Aarhus University Hospital, Denmark
We found aneuploidy in 4.7% of samples with fetal malformations, pathogenic CNV in additional 5.4% and variants of unknown significance (VOUS), likely pathogenic, in 1.6%. We have successfully replaced conventional karyotyping with CMA with a higher detection
rate, a shorter turn-around time in a clinically manageable way for both abnormal ultrasound and increased risk in a government financed system covering the central Denmark region of 1.2 million people.
13:40 Array-CGH (aCGH) as a First-Tier Test in All Invasive Prenatal Samples
Julian Nevado, Ph.D., Responsible for Genomics and Quality Manager, INGEMM, Spain
Given the increasing prevalence of NIPD testing for prenatal diagnosis, what is the best course of action with invasively-obtained prenatal samples? aCGH has been recommended by different experts and international guidelines as a first tier-test in many
postnatal clinical settings, but should this also apply prenatally? Based on our experience we will try to answer whether aCGH should be applied for all invasive samples and if it should be used as a first test or in parallel to Karyotype. Current
data in our lab, we will help to decipher the best option.
14:10 Refreshment Break
15:00 Polymorphism in the Promoter Region of Placental Protein 13/LGALS13 Gene as a Biomarker for Pre-Eclampsia
Hamutal Meiri, Ph.D., MBA, Coordinator of International Research, ASPRE Consortium, Israel
Gene polymorphisms are related to resilience/susceptibility to acquire various medical disorders including preeclampsia (PE). Our aim was to assess DNA polymorphisms in the LGALS13 sequence, encoding for placental protein 13 (PP13), in predicting PE,
given PP13’s unique placental expression and its potential role as a PE biomarker. It opens a new avenue for DNA-based prediction of a major pregnancy disorder related to placental dysfunction.
15:30 Risk Factors and Proteomic Biomarkers for Pre-Term Birth
A. Lopez Bernal, M.D., Ph.D., School of Clinical Sciences, Obstetrics
and Gynecology, University of Bristol, United Kingdom
The mechanism responsible for the onset of human labour remains a mystery but labour and delivery provoke changes in maternal plasma proteins associated with immune and defence responses and with lipid transport. We have used Tandem Mass Tag (TMT)
labelling for a comprehensive analysis of proteins in the maternal circulation, as well as with intrauterine tissues. Prostaglandins (PGs) have been implicated in parturition for many decades. We propose that reduced expression of certain biomarkers
as a potential mechanism through which smoking may contribute to preterm labour. Proteomic biomarkers will provide valuable insight into the mechanism of human labour and inform further research to improve the prediction and management of preterm
birth.
16:00 ROUNDTABLE BREAKOUT DISCUSSIONS
Topics include:
- Microarrays and Sequencing for Invasively-Obtained Samples
- Ethical and Genetic Counseling Issues for Prenatal Testing
- Developing Biomarkers for Preeclampsia and Pre-Term Birth Risk
- In-House vs. Service-Based Cell-Free DNA testing
- Prenatal Testing for Sub-Chromosomal Abnormalities
- Commercialization Potential for Fetal Cell-Based Non-Invasive Testing
17:15 Close of Day One
TUESDAY, 11 APRIL
08:00 Registration and Morning Coffee
09:00 Chairperson’s Remarks
09:05 Mosaic Whole Chromosome Uniparental/Biparental Disomy: A Common Feature in Trisomic Placentas?
Diane Van Opstal, Ph.D., Laboratory Specialist in Clinical Genetics, Erasmus Medical
Center, The Netherlands
The results of SNP array investigations of first trimester chorionic villi and term placentas, for confirmation of abnormal NIPT results, reflect the cytogenetic chaos that has been described in cleavage-stage embryos. Apart from confined placental
mosaicism involving trisomies and different structural chromosome aberrations, we also found mosaic whole chromosome uniparental disomy (UPD)/biparental disomy (BPD) which is rarely described in the literature, but which seems to be common in
trisomic placentas. Interesting cases will be presented.
09:35 Lessons from More Than 6000 NIPS from a Single Center, Including Use of NIPS in Complex Obstetrical Situations and Ultrasound Anomalies
Francois Jacquemard, Ph.D., Gynecology & Obstetrics, American Hospital
of Paris, France
From January 2013, more than 6000 NIPS have been processed through the Prenatal Diagnosis Center of the American Hospital of Paris (France). Details related to indications for testing and clinical finds will be reported, including rates for PPV, sensitivity
and false-positives. Lessons learned from these results, including that NIPS was also useful in some complex obstetrical situations when the risk of miscarriage or premature delivery was very high and contraindicated any invasive testing, allowing
almost full exclusion of main aneuploidies.
10:05 Expediting
NIPT Assay Development, Validation and Routine Monitoring
Ram Santhanam, MS, MBA, Director, Market Development - Reproductive Health, SeraCare Life
Sciences
As non-invasive prenatal testing (NIPT) becomes more mainstream, there is a critical need for patient-like reference materials to expedite new assay development, validation, external quality assurance (EQA) testing, and routine monitoring. We will
discuss case studies and performance data from a cross section of laboratories successfully utilizing these reference materials.
10:20 cfDNA Testing for 22q11.2 Deletion Syndrome Using a Targeted
Microarray Based Technology
Maximilian Schmid, M.D., Senior Director of Medical Affairs, Medical Affairs,
Roche Sequencing Solutions, United States
Deletion of 22q11.2 is the second most common cause of developmental delay and major congenital heart disease after Down syndrome1. The Harmony® Prenatal Test, used for assessing the probability of Down syndrome (Trisomy 21) and other chromosomal
conditions, has extended its menu to include 22q11.2 deletion testing. Top-line results of an analytical validation will be presented.
1. Rauch et al. Am J Med Genet A. 2006 Oct 1;140(19):2063-74
10:35 Coffee Break in the Exhibit Hall with Poster Viewing
11:15 Frequency of Fetal Submicroscopic Chromosome Aberrations in Low-Risk Pregnancies – A Systematic Review of Literature
Malgorzata Srebniak, Ph.D., Laboratory Specialist in Clinical Genetics,
Clinical Genetics, Erasmus Medical Center, The Netherlands
A systematic literature search was performed in order to establish the frequency of pathogenic submicroscopic copy number variants (CNVs) in fetuses without ultrasound anomalies and therefore no increased risk for unbalanced structural aberrations.
The aim was to determine the clinical value of high resolution fetal testing in the general pregnant population. We will show combined data of the relevant studies indicating the background risk for pathogenic submicroscopic aberrations in ca.
10,000 fetuses without increased a priori risk.
11:45 Clinical Implications of Testing for Sub-Chromosomal Abnormalities with NIPT
Yaron Yuval, M.D., Director, Prenatal Genetic Diagnostic Unit, Tel Aviv Sourasky Medical Center,
Israel
Detection rates for microdeletions may vary considerably depending on deletion size and positive predictive values (PPVs) - strongly influenced by prevalence - are expected to be low. Whole genome NIPS has recently been introduced, claiming the ability
to detect copy number changes throughout the genome. The benefits and limitations of expanded NIPS will be discussed with the aim of assisting clinicians and policymakers in incorporating NIPS into clinical practice.
12:15 Enjoy Lunch on Your Own
13:45 Chairperson’s Remarks
13:50 Non-Invasive Prenatal Testing of Monogenic Disorders by Droplet Digital PCR
Yves Rozenholc, Ph.D., Department of Statistics, University of Paris Descartes, France
Recently the use of NGS and digital-PCR using cell-free fetal DNA has been the basis for non-invasive prenatal tests for single-gene disorders. However, to date, it has mainly been offered for paternal and de novo mutation exclusion, the study of the maternal inheritance remaining challenging because of the high levels of maternal background. The development of a digital droplet PCR Uniformly Most Powerful Likelihood Ratio Test has been developed for a
range of different single gene disorders. Results using this approach, and recommendations related to blood volume to collect, for each type of single-gene disorder, will be discussed.
14:20 Toward Refined Non-Invasive Prenatal Testing by Digital Droplet PCR and Free Fetal DNA Enrichment
Neil Avent, Ph.D., Molecular Diagnostic and Transfusion Medicine, School of Biomedical &
Healthcare Sciences, Plymouth University, United Kingdom
Digital droplet PCR (ddPCR) is a significant enhancement over quantitative real-time PCR, and previous hardware associated with performing digital PCR. It has greater sensitivity and specificity than previously described methods, and has yet to
impact fully on its application in non-invasive prenatal testing. We have performed ddPCR on maternal plasma samples to detect paternally inherited genes (RHD and SRY), and have proved its superiority over real-time methods. By pre-enrichment
of maternal plasma samples this technique could readily be applied to the assessment of aneuploidy, providing a cheaper and more straightforward approach than current next-generation sequencing dependent methods to define aneuploidy.
14:50 Developing and Delivering a Non-Invasive Prenatal Diagnostic Service for Monogenic Disorders
Lucy Jenkins, FRCPath, Lab Director, Great Ormand Street Hospital, London, United Kingdom
The development of NIPD for single gene disorders, and particularly the impact of next-generation sequencing technologies that have been used to extend our service repertoire, will be presented. Previously limited to de novo mutations or paternal exclusion testing, we have extended this approach to the implement bespoke test development for families with rare disease. NGS has now facilitated relative haplotype dosage analysis (RHDO) for the diagnosis of recessive
conditions, even where parents carry the same mutation. Uptake of testing in the UK will be discussed along with issues that arise during NIPD delivery.
15:20 FEATURED POSTER: Looking to the Experience Obtained from Implementing Non-Invasive Prenatal Testing of Common Chromosomal Aneuploidies Using Cell-Free Fetal DNA in Iran
Mohammad Keramatipour, Ph.D., Assistant Professor, Genetics, Tehran University of Medical Science, Iran
15:35 FEATURED POSTER: Maternal Aneuploidy as a Cause of False Positive Noninvasive Prenatal DNA Screening Results
Jekaterina Shubina, Junior Researcher, Research Center for Obstetrics, Gynecology, & Perinatology, Russia
15:50 Refreshment Break in the Exhibit Hall with Poster Viewing
16:30 Panel Discussion with Cell-Free DNA Assay Providers
Michael Lutz, CEO, Life Codexx AG, Germany
Samantha Leonard, International Medical Director, Natera, France
Stephen Little, Ph.D., Chief Executive Officer, Premaitha Health, United
Kingdom
Daniel Grosu, M.D., MBA, Consultant, Integrated Genetics, United States
Gautam Kollu, Head of Market Development, RGH Market, Illumina Inc., United States
Maximilian Schmid, M.D., Senior Director of Medical Affairs,
Medical Affairs, Roche Sequencing Solutions, United States
18:00 Welcome Reception in the Exhibit Hall with Poster Viewing
19:00 Close of Day Two
WEDNESDAY, 12 APRIL
08:00 Registration and Morning Coffee
08:30 Chairperson’s Remarks
Patrizia Paterlini-Brechot, Ph.D., Cellular & Molecular Biology, University of
Paris Descartes, France
08:35 The Case for Cell-Based NIPT Using DNA from Fetal Cells
Ripudaman Singh, Ph.D., COO, ARCEDI Biotech Aps, Denmark
In our previous studies we have shown that extravillous trophoblasts enriched from maternal blood using specific markers can be used to detect chromosomal and sub-chromosomal variations in the fetal genome. We present data from our clinical
validation study which brings us a step closer to pursuing cell-based NIPT on all pregnancies, avoiding the use of invasive procedures.
09:05 Updated Results for Isolation and Genetic Characterization of Trophoblastic Cells for Prenatal Diagnosis of Genetic Diseases
Patrizia Paterlini-Brechot, Ph.D., Cellular
& Molecular Biology, University of Paris Descartes, France
The development of a non-invasive alternative to the invasive approaches for prenatal diagnosis of genetic diseases requires the use of fetal DNA non-mixed with maternal DNA, thus the availability of cells carrying the fetal DNA. We have
developed ISET, allowing the consistent recovery of trophoblastic cells from blood and cervical samples, and have developed methods for the extensive genetic analyses of the collected cells. We will present results of the use of trophoblastic
cells isolated by ISET for non-invasive prenatal diagnosis of trisomy 21 and monogenic disorders and discuss the technical issues involved in bringing to the market this new approach.
09:35 Advances in the Isolation of Fetal Cells from Maternal Blood
Leonard Kellner, President, KellBenx, United States
The isolation of fetal nucleated red blood cells (fnRBC) in maternal blood allows for a non-invasive diagnostic test of the fetal genome. Methods to analyze these cells will be presented and discussed.
10:05 FEATURED POSTER: Performance of Four Modern Whole Genome Amplification Methods for Copy Number Variant Detection in Single Cells
Lieselot Deleye, Ph.D. Student, Pharmaceutical Sciences, Ghent University
10:35 Coffee Break in the Exhibit Hall with Poster Viewing
11:05 Closing Panel Discussion: Predicting the Landscape for Prenatal Molecular Diagnostics over the Next Few Years
Panelists to be Announced
12:05 Close of Advances in Prenatal Molecular Diagnostics Conference