1 The 10th Santorini conference
After a 2-year hiatus, due to the recent COVID-19 pandemic, the 10th biannual conference
on Systems Medicine, Personalised Health and Therapy, under the auspices of the Santorini
Conferences Association (SCs), the International Federation of Clinical Chemistry
and Laboratory Medicine (IFCC), the European Federation of Clinical Chemistry and
Laboratory Medicine (EFLM), the Hellenic Society of Pharmacogenomics and personalized
Diagnosis and Therapy (EEPHARM), and the European Society of Pharmacogenomics and
Personalised Therapy (ESPT), took place in Santorini, Greece, between 23 and 26 of
May 2022.
It was sponsored by several companies: Randox (Crumlin, United Kingdom), Agena Bioscience
(Hamburg, Germany) and Cellecta as Gold Sponsors; Thermofisher Scientific (San Francisco,
United States) and PharmGenetix (Vienna, Austria), as Silver Sponsors; HMG systems
Engineering (Fuerth, Germany) as Bronze Sponsors, reinforced by specific supports
from the Santorini Conferences Association (SCs), The Austrian Society for Laboratory
Medicine & Clinical Chemistry (OGLMKC), Transgene (Strasbourg, France), The Journal
“Frontiers in Genetics” and strengthened with scientific meetings supported by the
University of Lorraine and INSERM (“Cercle Gutenberg”).
For this 20th anniversary, we attracted many world-renowned delegates (researchers,
scientists, biologists, pathologists, oncologists, genetic epidemiologists, pharmacogeneticists
and biobanking experts, both from the academia and the industry) from 28 countries.
The event was particularly successful and of high scientific quality. The 10th Santorini
Conference, under the Presidency of Sofia Siest, the director of the EA_1122; IGE-PCV
(http://ige-pcv.univ-lorraine.fr/en/), University of Lorraine, France, offered a diverse
and innovative scientific program, showcasing the work of 35 worldwide distinguished
speakers, who shared recent advances on personalized medicine with 127 attendees in
eight distinct sessions.
The Conference was initiated by a keynote round table on “PGx analysis in the medical
diagnostic laboratory–from science to clinical decision support (CDS)” and was followed
by a keynote lecture on “Advances in Cancer detection”.
The second day was focused on: “Liquid Biopsy, Past, Present, Future”—“Advances on
Cellular and Multi-Omic Approaches” and “Approaches for the Discovery of Drug Targets,
Resistance Mechanisms and Biomarkers” and included a flash oral communication session.
The third day sessions were on “Heart inflammation”, “Econ-Omics: Better Care for
Better Cost”, “Genetic Screening & Clinical Applications (part I and II.
The fourth and last day ended out with an oral communication session, a session on
“Digital Health” and a closing presentation made by Sofia Siest.
In this article we briefly outline the presentations delivered during the conference
and review the key messages and conclusions.
2 The odyssey from hope to practice
The conference was officially initiated by Sofia Siest, the President of the Santorini
Conference series, thanked all the members of the different committees and the sponsors.
She presented an overview of the outcomes of the previous conference and announced
the inauguration of the Santorini Conferences series under the umbrella of the Santorini
Conferences Association (SCs) and the development of the new website: www.santoriniconference.org.
She then underlined that this was the 20th Anniversary of the conference’s series
and outlined the content of the scientific program (presentations and speakers) and
the different topics to be covered.
Following the welcome session, Raute Sunder-Plassmann (Vienna, Austria) and Markus
Paulmichl (Salzburg, Austria) introduced the keynote round table, supported by the
“OGLMKC” and focused on Pharmacogenetics (PGx) Analysis in the Medical Diagnostic
Laboratory–From Science to Clinical Decision Support (CDS).
The major topics/highlights of the round table discussion included.
• Whole genome sequencing provides opportunities to identify new genetic factors for
efficacy and safety phenotypes or for explaining the missing phenotype heritability
seen in twin studies. New genetic variants could act as regulators of pharmacogenes
expression.
• Long read sequencing allows to unravel complex gene loci.
• Large biobanks offer the opportunity to discover pharmacogenomic phenotypes.
• Value in expanding pharmacogenomic research to diverse ancestry groups.
• Significance of personalized prescription.
• Advances in genotyping technologies and a concomitant drop in the costs and turnaround
time facilitate multi-gene analyses and preemptive pharmacogenetic testing in medical
diagnostic laboratories.
• Value in switching to extended PGx panel diagnostics.
• Adhere to current guidelines on pharmacogenomic testing and reporting.
• Importance of recommendations for standardization of pharmacogenetic terminology
and test design.
• Importance of providing clear, concise, and interpretable reports, providing details
on the test itself and the identified variants in a separate supplementary document.
• Need for integrating PGx data into the patient’s electronic health record and—if
available—in a Clinical Decision Support System (CDSS).
M. Ingelmann Sundberg (Stockholm, Sweden) described how the ability to interrogate
the whole genome provides us with unparalleled opportunities to identify new genetic
predisposing factors in an unbiased manner for both efficacy and safety phenotypes,
often leading to new insights into gene function and explaining the missing heritability
seen in twin studies. Rare variants, which are unaccounted for during standard genotyping,
actually explain up to 4%–6% of the variability in certain genes encoding enzymes
and transporters. Similarly, haplotypes in linkage equilibrium to variants defining
a specific CYP allele may cause altered CYP activity due to additional variants affecting
gene expression. Additionally, variants in genes not directly linked to the ADME gene
in question may influence its regulation, as recently demonstrated for the nuclear
factor 1B (NFIB)- dependent regulation of CYP2D6 expression and risperidone metabolism
in psychiatric patients. Long read sequencing technologies allow to unravel complex
gene loci such as the CYD2Dlocus.
M. Pirmohammed (Liverpool, United Kingdom) explained how the availability of large-scale
biobanks, such as the United Kingdom biobank, enables us to identify novel pharmacogenomic
phenotypes—but may be limited by the depth of clinical phenotypes within the biobanks.
Additionally, it is important to expand pharmacogenomic research to different ethnic
groups to ensure that what we discover and implement is relevant to all population
and that we do not exacerbate race and health inequalities.
Ron HN van Schaik (Rotterdam, Netherlands) presented how selected centers reported
good results for improving medication safety and efficacy by including common genetic
variations in genes encoding enzymes, transporters and targets into pharmacogenetic
guided treatment decisions. However, comprehensive and/or preemptive pharmacogenetic
analyses and their implementation into daily clinical practice is still not very common.
Physicians usually rely on reactively ordered PGx tests for selected variants in a
single or only a few genes and particularly on a short turnaround time to initiate,
adjust or change a standard therapeutic regime. Due to recent advances in genotyping
technologies and a concomitant drop in the costs and turnaround time for multi-gene
analyses, PGx panel diagnostics using single-nucleotide polymorphism (SNP) genotyping
arrays or next-generation sequencing (NGS) based approaches are established in a few
medical diagnostic laboratories. But further efforts of wider implementation to the
entire healthcare systems is needed. There is also a patient driven demand for PGx
guided treatment decisions and multigene/multivariant testing. Preemptive genotyping
would ensure that the appropriate genetic information to guide drug therapy is already
available when needed and will maximize the effect of PGx.
R. Sunder-Plassmann (Vienna, Austria) described how the availability of an abundance
of PGx data still represents a huge challenge for physicians, who frequently struggle
with the interpretation and application of PGx test results. Hence, PGx reports have
to be designed in such a way that the data is easy to comprehend, and the focus is
on the current medication and relevant actionable genetic variants, preferentially
in a separate short report for immediate use. In addition, a comprehensive report
explaining the patient’s pharmacogenetic profile in more detail should be also provided.
According to current recommendations (ACMG), the patient’s genotype, predicted PGx-phenotypes,
commonly used drugs that may be affected by the identified genotype, a statement that
alternative medication might be considered (if applicable) and resources for guidelines
should be reported. Additionally, they suggest reminding physicians that the accuracy
of the anticipated PGx phenotype is dependent on the variants identified and that
co-medication and drug-drug interactions may also influence the phenotype.
For future therapeutic decisions, PGx data should be included in the patients’ electronic
health records. Currently, only few in vitro diagnostic tests are commercially available
for PGx analyses. The majority of medical diagnostic laboratories that offer PGx panel
diagnostics use lab developed assays, which may vary in gene selection, variants detected,
and nomenclature for phenotype description. This may lead to discrepancies in the
test results between laboratories and the need for standardizing the minimal requirements
for a diagnostic PGx test.
Markus Paulmichl (Salzburg, Austria) closed the session with a presentation on “Standardization
in PGx diagnostics”.
George Dagher (Paris, France) and Sofia Siest (Nancy, France) introduced the keynote
lecture on “Advances in Cancer Detection” presented by Nickolas Papadopoulos (Baltimore,
United States). Nickolas Papadopoulos highlighted that the earlier a cancer is detected
the higher the chance for a successful outcome. For many cancers there are not any
screening modalities available. The ability to identify cancers through blood testing
is one of the most exciting advances in cancer diagnostics. In a screening setting
it provides the opportunity to detect multiple cancer types with a single test. Liquid
biopsy also has the potential to detect early signs of minimal residual disease and
recurrence. The presentation outlined these opportunities along with challenges associated
with such clinical applications. We discussed the biomarkers, technologies and the
type of studies required to develop and evaluate the utility of such tests, in the
first session of the next conference day.
3 Conference sessions
3.1 Session 1: Liquid BIOPSY, past, Present, future
The session was chaired by Georges Weryha, Nancy, France, and Georges Dagher, Paris,
France, and sponsored by AGENA.
Klaus Pantel (Hamburg, Germany) started the session with a talk on “Liquid Biopsy:
From Discovery to Clinical Implementation”, highlighting the utility of liquid biopsy
in clinical practice. The molecular analysis of circulating cell-free tumour DNA (ctDNA)
and circulating tumour cells (CTCs) released into the blood can provide clinically
relevant information as “liquid biopsy” (Pantel and Alix-Panabieres, 2019; Alix-Panabieres
and Pantel, 2021) and provide new insights into tumour biology (Keller and Pantel,
2019). A variety of targeted and non-targeted approaches have been used to assess
ctDNA, including NGS and MassARRAY-Based ctDNA assays (Belloum et al., 2020; Schneegans
et al., 2020). Liquid biopsy analyses with validated platforms provide information
on early detection of cancer, identification of cancer patients at risk to develop
relapse (prognosis), and it may serve to monitor tumour evolution, therapeutic targets
or mechanisms of resistance on metastatic cells. New promising liquid biopsy markers
include extracellular vesicles, circulating microRNAs and tumour-educated platelets
as well as circulating host cells. Technical standardization and clinical validation
of liquid biopsy assays are essential (Connors et al., 2020).
Catherine Alix-Panabières (Montpellier, France), continued the session with a talk
on the “Metastasis-Competent Circulating Tumor Cells in Colon Cancer”, elaborating
on the need of developing a reliable, standardized and robust method to expand CTCs
from different cancer types. Real-Time Liquid Biopsy has been introduced (Pantel and
Alix-Panabieres, 2010) as a new diagnostic concept predicated on the analysis of circulating
tumor cells (CTCs) or circulating tumor-derived factors, in particular, cell-free
tumor DNA (ctDNA). Highly sensitive liquid biopsy assays have been developed that
can now be applied to detect and characterize minimal residual disease (Pantel and
Alix-Panabieres, 2019; Alix-Panabieres and Pantel, 2021). Furthermore, CTCs are promising
new biomarkers for prognostic prediction and monitoring of therapies in patients with
solid tumors, as well as understanding the biology of metastasis in cancer patients
(Alix-Panabieres and Pantel, 2014). However, an in-depth investigation of CTCs is
hampered by the very low number of these cells, especially in the blood of colorectal
cancer patients. Thus, the establishment of cell cultures and permanent cell lines
from CTCs has become the most challenging task over the past year (Cortes-Hernandez
et al., 2020). Alix-Panabieres et al. described, in 2015, the in vitro expansion of
colon CTCs and established the first permanent cell line from CTCs of a metastatic
colon cancer patient (Cayrefourcq et al., 2015). This colon CTC line designated CTC-MCC-41
is in culture for more than 6 years and has been characterized at the genome, transcriptome,
proteome and secretome levels. This thorough analysis showed that CTC-MCC-41 cells
resemble characteristics of the original tumor cells in the colon cancer patient and
display a stable phenotype. The molecular portrait of CTC-MCC-41 line displays a very
specific transcription program completely different than those of the primary and
metastatic colon cancer cell lines (Alix-Panabieres et al., 2017). More recently,
Alix-Panabieres’ team characterized eight additional CTC lines using blood samples
from the same metastatic cancer: a unique biological material collected before and
after chemotherapy and targeted therapy, and during cancer progression (Soler et al.,
2018). More recently, they showed that the PI3K/AKT/mTOR signaling pathway plays a
key role in the proliferation of the CTC-MCC-41 line (Smit et al., 2020) and that
the selective treatment pressure in colon cancer drives the molecular profile of resistant
CTC clones (Cayrefourcq et al., 2021). Although, viable CTCs are not exploitable in
all patients with cancer, they are a precious tool to unravel the mechanisms of metastasis
formation and cancer cell dissemination through the identification/characterization
of the aggressive tumor cells that need to be eradicated. We are progressing very
fast in the field of liquid biopsy in cancer research. However, much effort should
now focus on developing a reliable, standardized and robust method to expand CTCs
from different cancer types (Alix-Panabieres, 2020). The establishment of CTC lines
represents a new opportunity to decipher the metastatic cascade and, hopefully, to
find ways to stop cancer dissemination.
Ed Schuuring (Groningen, Netherlands), took the floor and gave a talk on the “Detection
of clinically actionable mutations in NSCLC: is there a one-fits-all cell-free DNA
test for routine clinical practice?”. Circulating tumor DNA (ctDNA) is a potential
minimally invasive molecular tool to guide treatment decision making and disease monitoring
especially when no appropriate tumor tissue biopsy is available. A suitable diagnostic-grade
platform is required for the detection of tumor-specific mutations in circulating
cell-free DNA (ccfDNA) with high sensitivity. Dr Schuuring described the objective
of their study to investigate if a one-fits-all ccfDNA test exists for the different
applications in the molecular diagnostics of lung cancer such as molecular profiling
for treatment-decision-making, response monitoring and detection of treatment-resistant
mechanisms. The design of the study was to determine the concordance between various
ccfDNA tests using plasma collected at various time-points during therapy from NSCLC
patients treated with tyrosine-kinase and immune checkpoint inhibitors. The researchers
evaluated the Roche Cobas® EGFR Mutation Test v2, the Agena UltraSEEK® Lung Panel,
tumor-mutation-specific Bio-Rad®-ddPCR assays and the Roche AVENIO ctDNA Expanded
NGS Kit. Based on the results presented, the concordance to detect clinically relevant
mutations in plasma comparing the different ccfDNA tests was >90% for those mutations
covered by the different assays. The concordance between therapeutically targetable
mutations detected in tumor tissue with NGS and in the pre-treatment plasma samples
was high for all assays (∼80%) and in agreement with reported data. In conclusion,
there seems to be no one-fits-all ccfDNA test for all clinical application. To select
the appropriate ccfDNA test for clinical questions that lead to actionable mutations,
aspects like the complexity of the test, costs, reimbursement issues, turn-around-time,
the number of relevant mutations covered by each assay, expertise of the diagnostic
lab and availability of ccfDNA, are important. Several studies presented were supported
by the CANCER-ID consortium (including Roche and Agena Bioscience) and unrestricted
research grants of Bristol Myers Squibb, Bio-Rad, Biocartis, and Agena Bioscience.
Various studies were based on collaborations with MUG Graz (E Heitzer), Imagenome
Montpellier (P-J Lamy) and UKE Hamburg (H Wikman).
The session was concluded by the talk from Oellerich et al. (Göttingen, Germany),
on “Donor-derived cell-free DNA testing in organ transplantation: a value proposition”.
Dr Oellerich outlined a value proposition for donor-derived cell-free DNA (dd-cfDNA)
testing in organ transplantation. There is a need to improve personalized immunosuppression
in organ transplantation to reduce premature graft loss. Biomarkers are needed to
better detect rejection, asymptomatic graft injury, and under-immunosuppression. Assessment
of minimal necessary exposure to guide tapering and prevent immune activation is also
important. There is robust clinical evidence from more than 50 published studies supporting
the role of dd-cfDNA for monitoring graft integrity and detection or exclusion of
rejection. The value proposition for the patient includes earlier transplant injury
intervention, less full blown rejection risk, an alternative to invasive biopsies,
personalized immunosuppression with potential for improved long-term outcome. Transplant
physicians benefit from better immunosuppressive guidance and having an alternative
when biopsies are refused or contraindicated. Further advantages are improved biopsy
interpretation, less trial and error changes in immunosuppression, and less time dealing
with complications. The laboratory medicine specialist can provide more effective
services. Hospital management and insurance companies could benefit from more cost-effective
surveillance of transplant recipients. Potential cost savings would be due to fewer
biopsies as a result of the high negative predictive value, fewer retransplantations,
or less organ failure with return to dialysis. A pathway to implementation and metrics
was suggested to measure the effectiveness of dd-cfDNA testing.
3.2 Flash Communications session (supplement)
The session was chaired by Stavroula Kanoni, London, United Kingdom and Vangelis Manolopoulos,
Alexandroupolis, Greece
3.3 Session II–advances on cellular and multi-omic approaches
The session was chaired by Behrooz Z. Alizadeh, Groningen, Netherlands and Georges
Weryha, Nancy, France
The first talk of this session was from Colin J.H. Brenan (Boston, United States),
presenting the “Engineering a Rational Approach to Precision Oncology Dugs”. Clinical
trials of new oncology drugs have a staggering 97% failure rate typically due to toxicity
or lack of drug efficacy. One common issue is failure to understand the mechanism
of action and misidentification of putative biomarkers indicative of drug response.
To address this issue, Dr Brenan and his team developed and clinically deployed an
innovative implantable microdevice (called the Nanonail™) for functional and simultaneous
intra tumor molecular profiling of tumor sensitivity to up to 18 different oncology
drugs and/or drug combinations per microdevice at the single cell level and with the
tumor in its native microenvironment. Each unique drug or drug combination was loaded
into one of the micromachined depots of the Nanonail for delivery of a controlled
dose of drug to the surrounding tumor tissue once inserted into the tumor with a standard
fine needle biopsy tool. After 1–3 days the device was recovered with a plug of surrounding
tissue and processed according to standard histopathology protocols. Thin sections
of the zone of drug-tumor interaction were individually analyzed to create a detailed,
high resolution spatial multi-omic profile of the tumor response to each agent. An
additional benefit, molecular profiling the tumor-drug response along the spatial
diffusion profile of drug from each depot provides key information on tumor response
to different drug doses. Dr Brenan provided examples on how application of single
cell functional profiling can reveal novel, potent anti-tumor drug combinations and
in particular the combination of an immunotherapy with molecularly targeted or cytotoxic
agents.
The second presenter of this session was Ekaterini Chatzaki (Alexandroupoli, Greece),
describing her recent work on “Biomarker discovery in the era of automated machine
learning: from targeted to data-driven approaches”. Biomarkers are the cornerstone
of precision medicine: identified as a measurable indicator of some biological state
or condition, they promise to offer solutions for accurate diagnosis, prognosis and
therapeutic monitoring. Dr Chatzaki and her team have been studying methylation in
liquid biopsy material in different pathological conditions such as cancer and diabetes.
The team has moved gradually from hypothesis-driven to (big) data-driven approaches,
as modern -omics technologies lead the accumulation of large precious multi-parametric
biological datasets. They employed ad hoc auto machine-learningtools for data extrapolation,
delivering low-feature validated models/classifiers and suggest that this approach
can have unprecedented added value in different medical conditions.
3.4 Session III–approaches for the discovery of drug targets, resistance mechanisms,
and biomarkers
The session was chaired by Stavroula Kanoni, London, United Kingdom and Behrooz Z.
Alizadeh, Groningen, Netherlands and sponsored by CELLECTA
Paul Diehl (Cellecta, Inc, Mountain View, California, United States) presented “Flexible
and Scalable Genetic Screens for Discovery and Characterization of Novel Therapeutic
Targets”. The measurements of changes in gene activation and expression provide a
basis to understand the genetic changes that cause biological responses of interest.
Cell-to-cell gene disruption induced by CRISPR and other gene-perturbation technologies
help tease out the drivers required for these responses. Dr Diehl discussed how adaptations
of these two screening approaches can be used to discover the genetic drivers responsible
for phenotypic variabilities, such as drug sensitivities, disease variation, and degrees
of differentiation within cell populations, across tissue microenvironments, and between
single cells. Also from the same company, Alex Chenchik talked about “Immunophenotyping
of T Cell receptor and B Cell receptor clonotypes”. T Cell receptor (TCR)/B Cell receptor
(BCR) repertoire profiling holds great potential for understanding disease mechanisms.
Dr Chenchik explained how they introduced a novel technology for profiling of all
human TCR and BCR variable regions and phenotypic characterization of immune cells
in bulk and at the single-cell level in PBMCs and immune cell fractions. Preliminary
data showed that TCR/BCR clonotype analysis combined with targeted expression profiling
of immune cells can be applied for large-scale discovery in several immune-responsive
model systems.
3.5 Session IV–heart inflammation
The session was chaired by Panagiotis Deloukas, London, United Kingdom and Georges
Weryha, Nancy, France and sponsored by RANDOX
Federica Marelli-Berg (London, United Kingdom) gave a talk on “Tracking T cell-mediated
autoimmunity in the heart”. Autoimmune cardiac inflammation is becoming recognized
as a key contributing factor in heart muscle diseases. Despite advances, the functional
features of cardiac immunity in humans remain largely undefined, due to the technical
challenges of studying the immune response in-situ. Dr Marelli-Berg and others described
a population of cardiotropic T Cells (cT-cells) characterized by the expression of
the hepatocyte growth factor receptor cMet and the chemokine receptors CCR4 and CXCR3.
They showed that memory, activated cT-cells significantly increase in the circulation
and in the heart of patients with inflammatory cardiomyopathies, but not in acute
myocardial infarction or healthy controls. cT-cells divide preferentially in response
to the autoantigen cardiac myosin and display similar functional features in acute
and chronic cardiac inflammation. In experimental autoimmune myocarditis, which recapitulates
the autoimmune phase of human myocarditis, development of cT-cells and disease can
be prevented by pharmacological cMet inhibition, suggesting a causative role for this
T Cell subset.
Next, Behrooz Z. Alizadeh (Groningen, Netherlands) described the “Predictive value
of Inflammatory causes of vascular disorders in Personalized medicine”. The pivotal
role of inflammation in cardiovascular diseases (CVD) has been scrutinized for a century.
Accumulating number of studies suggest the involvement of specific molecular pathways
in the disease mechanism, which are represented by inflammatory biomarkers and are
claimed potential targets for therapeutics (NFκB, OPN). However, there is little known
whether these associations are causal and are dependent on the dominant type of inflammatory
cells. Dr Alizadeh presented the latest results of their studies on the causal association
of inflammatory biomarkers with major CVD phenotypes, by using genetic risk. They
also evaluated the potential application of inflammatory biomarkers in better prediction
of disease outcomes. Future investigations should focus on the crosstalk between causal
inflammatory biomarkers, the type of inflammatory cell involved, in the pathological
contexts of cardiac cells and may eventually lead to specific inflammatory-based therapies
for the personalized prevention and treatment of CVD.
The last talk in this session was given by Helena Murray (Randox Laboratories Limited,
Crumlin, Co. Antrim, United Kingdom), discussing the “Development of a Type I Diabetes
Genetic Risk Array”. Differentiating between Type 1 diabetes (T1D) and Type 2 diabetes
(T2D) is challenging due to the increasing incidence of childhood obesity blurring
the traditional T1D versus T2D timelines. More young people are getting T2D and T1D
can occur at any stage in life and an increasing number of cases of T1D is also occurring
at old age. Currently available diagnostic tests have several limitations in accurately
diagnosing diabetes subtypes with up to 15% of young adults wrongly classified and
treated. The aim of this study was to consider genetic predisposition as an aid to
improve diabetes classification. Genetic predisposition to diabetes is largely determined
by the presence of human leukocyte antigen (HLA) genes. Genome-wide association studies
have identified additional non-HLA SNPs, robustly linked with T1D. Combining these,
a 10 SNP genetic risk score (GRS) was developed which can aid discrimination between
T1D and T2D, particularly when used in conjunction with clinical features and autoimmune
markers. The assay employs multiplex Polymerase Chain Reaction (PCR) coupled to Biochip
Array Technology (BAT, Randox Laboratories Ltd., Crumlin, UK) to genotype 10 SNPs
associated with T1D (Oram et al., 2016). Assay optimization and specificity was achieved
using pre-characterized DNA samples and initially validated by testing DNA samples
(n = 259) provided by University of Exeter. The T1D GRS array is capable of rapidly
detecting all 10 SNPs associated with T1D. Through an associated algorithm, the array
can generate a T1D Genetic Risk Score, which in conjunction with conventional methods,
can distinguish T1D from other subtypes. This assay has potential to prevent misdiagnosis
of diabetes and facilitate improved patient management.
3.6 Session V–econ-omics: Better care for better cost
The session was chaired by Georges Dagher, Paris, France and Belgin Süsleyici, Istanbul,
Turkey
The first talk in this session was given by one of the chairs, George Dagher (Paris,
France), on “Big data, Artificial Intelligence and ethics”. Big data is certainly
an essential component of digital science and technology and also of machine learning,
robotics, and new means of communication. The information that the data initially
contains, is considerably enriched by cross-referencing data. Highly diverse, this
data can be related to health or wellbeing. One of the characteristics of big data
in health is the blurring of the distinctions underpinning implementation of the ethical
principles that promote the protection of individual rights in health. Precise knowledge
of individuals and of their state of health creates a risk of profiling, which threatens
the protection of private life and may lead to stigmatization of people or groups.
Such stigmatization threatens private life, but also the principles of solidarity
and equity which are the basis of our health system. Care and business are becoming
increasingly hard to distinguish, as a result of the transformation of care and of
the healthcare market. The need for protection of the individual must be reaffirmed
and its modalities redefined, to dispel the threat of a society under the surveillance
and control of multiple providers acting for various purposes.
The second talk in this session was given by Uwe Oelmueller (QIAGEN GmbH, Hilden,
Germany) on “Standardized Preanalytics: The Key for Reliable Diagnostics, Research
and Biobanking”. Molecular in vitro diagnostics and research have allowed great progress
in medicine including diagnostics. However, profiles of these molecules (nucleic acids,
proteins, and metabolites) can change significantly during specimen collection, transport,
storage, and processing. This can make the outcome from diagnostics or research unreliable
or even impossible. High quality specimens with preserved analyte profiles are crucial
for reliable diagnostics, biomedical research and biobanking. Specifying, developing
and verifying pre-analytical workflow parameters for diagnostics tests has consequently
become a requirement by new European legislation. The EU SPIDIA Consortium (2008–2013)
developed new pre-analytical technologies for preserving molecular profiles in human
specimen and generated broad evidence that guidance to laboratories on pre-analytical
workflows improves analytical test results. Based on these results, the CEN/TC 140
for “in-vitro diagnostic medical devices” had released first 9 European Technical
Specifications for pre-analytical workflows addressing different blood, other body
fluids and tissue based molecular applications. In 2018 and 2019 they progressed to
International Standards at the ISO/TC 212 for “clinical laboratory testing and in
vitro diagnostic test systems”. The successor EU SPIDIA4P consortium project (2017–2021),
supported by a large international network, has broadened to a final portfolio of
22 pre-analytical CEN and ISO Standards intending to improve in vitro diagnostics
and biomedical research, has developed corresponding External Quality Assurance (EQA)
and is driving international implementation. The SPIDIA project received funding from
the EU’s FP7 under grant agreement no. 222916. The SPIDIA4P project received funding
from the EU’s Horizon 2020 research and innovation program under grant agreement no.
733112.
3.7 Session VI—first part–genetic screening and clinical applications
The session was chaired by Guillaume Paré, Hamilton Canada and George Dedoussis, Athens,
Greece
Georges V Dedoussis (Athens, Greece) presented recent work on the “Omics and Mastiha
treatment in NAFLD - The EU Mast4Health program”. Non-alcoholic fatty liver disease
(NAFLD) is a major public health concern in both industrialized and developing nations,
with an estimated global incidence of 25% in the general population and with limited
treatment approaches. The Mast4Health consortium investigated the effect of the nutraceutical
Mastiha supplement in the omics profile of patients with NAFLD, within a multicenter,
randomized, double-blinded, and placebo-controlled clinical trial design. Based on
the results, there was an improvement in liver inflammation and fibrosis (as assessed
by MRI and the use of the sensitive LiverMultiScan software). Post-treatment levels
of both Liver Inflammation Fibrosis score (LIF) and iron-corrected T1 (cT1) were lower
in the Mastiha group compared to the Placebo among volunteers with BMI>35 kg/m2. The
Bray-Curtis dissimilarity index between baseline and post-treatment bacterial communities
was larger in the Mastiha group versus the Placebo. The metabolomic analysis showed
a significant reduction of Lysophosphatidylcholines and Lysophosphatidylethanolamines
in the Mastiha group suggesting that Mastiha exhibits a beneficial effect in phospholipid
homeostasis. In conclusion, after 6 months of Mastiha supplementation, the investigators
observed a significant improvement on microbiota dysbiosis and lipid metabolite levels
in patients with NAFLD (Amerikanou et al., 2021). This project received funding from
the European Union’s Horizon 2020 research and innovation program MAST4HEALTH under
the Marie Skłodowska-Curie grant agreement no 691042. (NCT03135873, www.clinicaltrials.gov).
3.8 session VI—second part–genetic screening and clinical applications
The session was chaired by Guillaume Paré (Hamilton Canada) and Csilla Sipeky (Brussels,
Belgium).
The first speaker in this session was Panagiotis Deloukas (London, United Kingdom)
discussing “Polygenic Risk Scores, application and challenges in cardiovascular disease
prediction”. A total of 14% of adults above the age of 16 have doctor-diagnosed cardiovascular
disease (CVD) which is a leading cause of death (∼25% of all deaths) and disability
in the United Kingdom. Coronary Heart Disease (CHD) is the leading cause of CVD death
in both sexes (14% men, 8% women). South Asians have a 2-fold higher risk for CHD
compared to European-descent individuals and an earlier onset of disease. A polygenic
risk score (PRS) aggregates all known genetic variants associated to the disease.
Dr Deloukas and others developed a European CHD-PRS using the data from the latest
CARDIoGRAMplusC4D meta-analysis in 1,165,690 participants including 181,522 cases
(2.3 M genetic markers) and evaluated this new CHD-PRS in the Malmo Diet and Cancer
study, confirming its predictive value and ability to predict secondary cardiovascular
events. LDL-cholesterol is a major CHD risk factor. In a parallel study of LDL-cholesterol
in 1.65 M individuals, the team showed that the multi-ancestry PRS had the best or
near-best performance in each ethnic group tested, with improved or equivalent prediction
relative to ancestry-matched scores (Graham et al., 2021). The team is further developing
a multi-ancestry CHD PRS and aim to validate its performance in 49,000 British South
Asians. Whilst pursuing investigation of disease prevention in the British population,
the team is also assessing the cost-effectiveness and clinical value of the CHD-PRS
in improving management of those with established disease. As part of a national UK
effort (Our Future Health program) the aim is to look at disease severity and onset
in patients with higher PRS, including rare CVD.
The next speaker, Robert Barouki (Paris, France) gave a talk on “The relevance of
non-genomic stressors: deciphering environmental factors for the next decade”. Climate
change, urbanisation, chemical pollution, and disruption of ecosystems, including
biodiversity loss, affect our health and wellbeing. HERA is an EC-funded H2020 CSA
project aiming at providing a research agenda for the next 10 years in the field of
environment climate and health (https://www.heraresearcheu.eu/). The agenda identifies
six major research goals in these fields. These include research to 1) reduce the
effects of climate change and biodiversity loss on health and environment, 2) promote
healthy lives in cities and communities, 3) eliminate harmful chemical and physical
exposures, 4) improve health impact assessment and implementation research, 5) develop
infrastructures, technologies and human resources and 6) promote research on transformational
change towards sustainability. Numerous specific recommendations for research topics
are presented under each research goal. The results call for an unprecedented effort
to support a better understanding of the causes, interlinkages and impacts of environmental
stressors on health and the environment. This will require breakdown of silos within
policies, research, actors as well as in our institutional arrangements in order to
enable more holistic approaches and solutions to emerge.
Next in the session, Guillaume Paré (Hamilton, Ontario, Canada) presented his recent
work on “Digging deep for translational gold: Multi-omics approach to cardio-metabolic
traits”. Despite recent advances in acute diagnosis and treatment of cardio-metabolic
diseases (CMD), the development of new blood biomarkers for risk stratification has
been slow. The majority of reported biomarker-disease associations fail to enter clinical
practice due to their inabilities to discriminate risk, or more importantly, due to
a lack of evidence that they represent causal associations with risk of disease. Distinguishing
modifiable, causal mediators from the many biomarkers that are statistically linked
to CMDs is a primary challenge in molecular epidemiology. Truly causal biomarkers
such as LDL cholesterol have been invaluable in the prevention, treatment and identification
of at-risk individuals. Dr Paré proposed an integrated genomic-proteomic (biomarker)
approach to identify novel causal mediators of CMDs, and illustrated with examples
from coronary artery disease, stroke, diabetes, chronic kidney disease and obesity.
This approach is based on Mendelian Randomization (MR) that protects against confounding
and reverse causation. Integrating genetics and high-throughput proteomics holds the
promise of better risk stratification, identification of new disease pathways, and
paves the way for novel therapeutic interventions.
Next in this session, Stavroula Kanoni (London, United Kingdom) gave an overview of
recent developments around the “Genetic susceptibility for COVID-19 infection and
severity”. The COVID-19 pandemic, caused by infection with SARS-CoV-2, has led to
a total of 373 M cases worldwide and 5.5 M deaths. The SARS-CoV-2 infection has varied
consequences, ranging from asymptomatic, to mild flu-like symptoms, to life-threating
consequences like viral pneumonia and acute respiratory distress syndromes. Risk factors
associated with the disease severity include increasing age, male gender, other comorbidities,
and ethnicity. Furthermore, host genetic factors have also been identified as risk
factors of SARS-CoV-2 infection or severe consequences of COVID-19, through genome-wide
association studies (GWAS), whole-genome sequencing (WES) and candidate gene studies.
Genes implicated with infection susceptibility or disease severity are involved in
key pathophysiological processes, including viral entry into cells, immunity, and
inflammatory responses. The most putative causal COVID-19 genes include SLC6A20, ABO,
CXCR6, INFAR2, OAS (1,2,3), DPP9, TYK2, ACE, MUC5B and FOXP4 and are linked to increased
susceptibility and/or severity. Genetic predisposition to severe COVID-19 is also
associated with deep venous thrombosis, morbid obesity, renal failure, pulmonary heart
disease and respiratory failure. Large scale observational and Mendelian randomization
studies have identified smoking as a risk factor for infection and severity, while
there is no protective effect of vitamin D on COVID-19 susceptibility, severity, or
hospitalization. The development of vaccines against SARS-CoV-2 have proven very efficient
at halting the spread. The emergence of mutated variants of the virus are causing
concern but vaccine engineering could be implemented to match the need of a response
to the new variants. It is very crucial to increase the genomic surveillance around
the world and at the same time increase the vaccine uptake.
3.9 Selected abstracts—oral communications session (supplement)
Session chaired by Stavroula Kanoni, London, United Kingdom and Vesna Dimitrejevic
Sreckovic, Belgrade, Serbia
3.10 session VII–digital health
The session was chaired by Alexander Haliassos, Athens, Greece and Ekaterini Chatzaki,
Alexandroupolis, Greece
Alexander Haliassos (Athens, Greece) presented recent advances in “Mobile Health (mHealth)
and Internet of Things (IoT)”. When internet-based enabling technologies are coupled
with new capabilities in mobile communications providing remote access from everywhere
using web enabled smart personal devices (phones, tablets and laptops) an opportunity
is created that can revolutionize, not only the scope, but also the process healthcare.
mHealth (or m-health) is an abbreviation for mobile health, a term used for the practice
of medicine and public health supported by mobile devices such as: mobile communication
devices, mobile phones, tablets, PDAs, and wearable devices such as smart watches,
for health services, information, and data collection, information exchange and communication
via the Internet. mHealth is one aspect of eHealth that is pushing the limits of how
to acquire, transport, store, process, and secure the raw and processed data to deliver
meaningful results. mHealth started at the industrialized nations but emerged in recent
years as an application for developing countries, stemming from the rapid rise of
mobile phone penetration in low-income nations where they face a plethora of constraints
in their healthcare systems. It provides greater access to larger segments of a population
in developing countries and improves the capacity of health systems in such countries
to provide quality healthcare. But there are concerns about the accuracy and unregulated
status of health apps.
The Internet of Things concept has three pillars for its development:
Connectivity: The universalization of Internet allows everyone to connect with high
bandwidth through cellphones or wireless networks (WiFi).
Sensing Devices: Universalization of cheap devices with sensing capabilities. There
are sensors for any of the five human senses. Smartphones can sense in which position
they are, if the user is looking at them, if the user is speaking to them, how fast
the user is moving and where in the world it is placed. Recently are in development
more sophisticated sensors that can detect smells and flavors.
Computational Power: Not only today’s devices (like smartphones, tablets) have the
same computational power of IBM’s 80s computers of the size of a room, but any device
is able to access additional computational power through the Cloud and its virtual
services.
The digital divide describes the differences between those who have access to the
Internet and those who do not because of economic reasons. It describes also the lack
of computer competency or self-efficacy and/or the lack of communications infrastructure.
This situation can impact negatively the use of the above described technologies in
many areas of our world. This can be partially compensated using specialized applications
off-line on computers or mobile devices.
Ivan Brandslund (Odense, Denmark) continued the session with a talk on “AI in cancer,
emergency and COVID-19”. Artificial Intelligence (AI) is a promising technology to
use in analysis of numerous data and in diagnostics. Based on the assumption that
the effect of any disease would change the pattern of laboratory test results and
thus a possibility that analysis of these patterns could evaluate absolute risk for
specific diseases, Dr Brandlund et al. have tested the ability of AI to predict risk
of cancer in patients consulting their general practitioner, the predictive values
for sepsis, death and specific diseases in emergency received patients as well as
analyzed consecutive data from patients admitted with severe disease caused by COVID-19.
The results were that the risk score of cancer can be measured from 0% to 80%, emergency-received
patients outcome could be predicted with area under the curve in ROC analysis of between
87% and 92% and that the analytical test with the highest predictive value for outcome
in COVID-19 is the absolute concentration of virus particles in the blood at admittance.
AI is likely to be a part of the clinical diagnostic methodologies within the next
10 years.
3.11 Session VIII–pharmacogenomics and post-marketing applications
The session was chaired by Charity Nofziger, Salzburg, Austria and Vangelis Manolopoulos,
Alexandropoulis, Greece
The first speaker of this session, Ron HN van Schaik (Rotterdam, Netherlands), talked
about “Pharmacogenetics testing in a healthcare system: opportunities and challenges”.
For the most effective use of pharmacogenetic testing, the uptake in a healthcare
system is key. This stretches from easy and straightforward ways to ask for testing,
easy sample collection, trustable, accurate and timely results, but also reporting
and reimbursement issues. When in a country, several laboratories are performing the
testing, harmonization of testing, interpretation and reporting become essential topics.
In Netherlands, they managed to implement much of this chain, including that every
pharmacist in the country can provide genotype-based dosing advice for more than 100
drugs. To have the system operating, continuous education of specialist, GPs, pharmacists,
students, patients and insurers was organized. One challenge still to be addressed
in the country is the fact that genotype to phenotype interpretations may change over
time. It is important to have genotype portals, or national electronic health records
available to notify already genotyped patients about any changes in interpretation.
Although this brings another aspect forward: how does the patient react to this change
of interpretation when they were treated based on the original reports? This is an
interesting challenge that should be discussed within the scientific community.
Next, Charity Nofziger (PharmGenetix GmbH, Anif/Niederalm, Austria), presented “Efforts
of the Pharmacogene Variation Consortium (PharmVar) to facilitate the interpretation
of pharmacogenetic test results and guide precision medicine”. The Pharmacogene Variation
Consortium (PharmVar) is the home for pharmacogene nomenclature that serves as a centralized
data repository for single nucleotide variants (SNVs) in PGx related genes. Its main
goal is to catalogue allelic variation in genes that play a role in the metabolism,
disposition and response to drugs, and provide a unifying and standardized nomenclature
system for the entire PGx community. Dr Nofziger gave a quick overview of the website
and highlighted its main features, tools and resources to facilitate PGx related work.
Ferrier Le (Thermo Fisher Scientific, Santa Clara, United States) presented a “Method
for CYP2D6 Copy Number Variation Analysis using Multiplex Digital PCR on the QuantStudio
Absolute Q System”. The team developed a method using multiplex TaqMan® digital PCR
to analyze CYP2D6 copy number variation using the QuantStudio™ Absolute Q™ Digital
PCR System. The system’s four optical channels enabled the development of a custom
4-plex digital PCR assay for the copy number variation analysis of three CYP2D6 regions
(exon 9, intron 2, and 5′ UTR) in a single digital PCR reaction. The assay performance
was verified using a panel of reference DNA samples. Compared to existing methods,
the workflow using the Absolute Q system with the multiplex CYP2D6 assay reduced the
time to results for copy number variation analysis without compromising accuracy or
performance.
The session continued with Ingolf Cascorbi (Kiel, Germany), talking about “Pharmacogenomics
of tyrosine kinase inhibitor resistance”. Tyrosine kinase inhibitors (TKI) are widely
used in the modern treatment of malignancies, such as the treatment of chronic myeloid
leukemia (CML) through imatinib, inhibiting the catalytic domain of the BCR-ABL fusion
gene product. However, resistances may be caused by BCR-ABL independent mechanisms.
The group identified mechanisms of drug resistance applying CML in-vitro models of
imatinib as well as nilotinib resistance. They were able to demonstrate significant
upregulation of the ABCG2 efflux transporter with strong association to deregulation
of specific miRNA. These mechanisms could be reversed only under treatment-naive conditions.
Further investigations revealed substantial changes of gene expression and (epi)genetics,
related established oncogene signaling pathways but interestingly also to cell adhesion
pathways (Kaehler and Cascorbi, 2021).
The final talk in this session was by Belgin Süsleyici (Istanbul, Turkey) on “Precision
Medicine in Routine Turkish Clinical Practice: Now and in the Future”. In recent years,
traditional treatment plans have been augmented by precision medicine approaches.
However, there are still significant issues to be overcome in incorporating these
approaches into routine care and integrating new research data to clinical practice.
In collaboration with many clinicians and scientists, and with significant support
of the Ministry of Health in Turkey, the group started studies by performing pharmacogenetic
analyses according to the drug-use status of patients from cardiology, oncology, psychiatry
as well as physical therapy and rehabilitation clinics. The effects of CYP2C9 and
VKORC1 polymorphisms on warfarin-dose requirements in Turkish patients were determined
and as a conclusion to the CYP2C9 *2, *3, VKORC1 9041 G>A polymorphisms were found
to explain the considerable proportion of inter-individual variability in warfarin
dose requirement. The individualized metoprolol doses to be used in treatment of heart
rates and blood pressures for cardiac patients and tramadol responses for physical
therapy and rehabilitation patients were related to CYP2D6 genotyping, therefore pharmacogenetic
results of CYP2C6 are being considered before drug prescriptions. In oncology, the
relationships between 5-FU treatment-related adverse events and DPYD, MTHFR and TYMS
gene polymorphisms involved in 5-FU metabolism in colorectal cancer (CRC) patients
were evaluated. Together with genes affecting 5-FU response, the relationship between
bevacizumab use and specific VEGF polymorphisms, have been investigated based on the
survival time and metastasis in colorectal cancer patients. The results obtained from
the PDL-1 checkpoint inhibitor have been used to produce a real-time PCR kit that
is able to help clinicians determine the immunotherapeutics they will be using in
targeted therapies. Based on the assumption that the variations in endothelial-mesenchyme
transformation inducers like SNAI1 and LOXL2, may have synchronous effect on metastases
resulting with malignant phenotype, genotyping results are being recorded for these
gene polymorphisms for their clarification to be used in PGx routine. This data will
contribute to the understanding of both malignancy and the potential of new therapeutic
targets to be used in the treatment processes of various disease.
3.12 Posters
Twenty-five posters (Supplementary) were presented that were classified in two groups
(group 1: “-omics” biomarkers and group 2: Pharmacogenomics). Jaroslav Hubacek and
his team won the “Omics” poster award, granted by The Santorini Conferences Association
(SCs), for their work on the “Apolipoprotein L1 variability is associated with increased
risk of renal failure in the Czech population”. Päivi Hirvensalo and her team won
the “Pharmacogenetics” award, granted by the European Society of Pharmacogenomics
and Personalised Therapy (ESPT), for their work on “Pharmacogenomics of celiprolol”.
4 Conclusion
During the 10th Santorini Conference, we enjoyed high quality talks and poster presentations,
covering a range of recent advances in the area of personalized medicine. World-renowned
scientists discussed state of the art approaches of introducing PGx in the clinical
practice, the use of liquid biopsies in the prognosis of cancer, the use of biomarkers,
genetic and polygenic risk scores and multi-omics in the prediction of cardiometabolic
traits and the implementation of AI in different areas of research and patients’ treatments.
We are looking forward to our next conference scheduled for 21–24 May 2024.