In translational research the question arises how to prepare researchers for the requirements of clinical studies in a way that can facilitate the transition of knowledge from basic, preclinical research with animals to clinical studies with humans. The aim of the presented reverse engineering approach is to familiarize basic researchers with the requirements and characteristics of the clinical study culture and in this way to prepare the basic researcher to create a more efficient translational experimental process. The reverse engineering approach consists, anong other help and guidance, of a preclinical evaluation guide for animal models and requirements for cell and gene therapy products prior to start of clinical study; process descriptions including the stages from preclinical research to phase I transition process to be used for analysis of the number of animals and necessary important experimental data to be generated for cell and gene therapy studies; compilations of the necessary legal provisions and advice received from Paul-Ehrlich Institute in Germany and AFSSAPS in France; list of international requirements for animal models, costs of animals for studies, special requirements for biotherapy studies, applicable guidelines for preclinical studies, demands for new investigational medicinal products. The relevance of the animal model is a focus, especially the consideration that a similar response in human and animal cells in vitro, does not guarantee that the in vivo response is similar.

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W. Kuchinke (2009)
Improve the transition from preclinical to
clinical research by reverse engineering of
clinical trials
WOLFGANG KUCHINKE
UNIVERSITY DUESSELDORF, DUESSELDORF, GERMANY
GMDS 2009, ESSEN, GERMANY
SEPTEMBER 7, 2009

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Introduction
●
In clinical studies on regenerative therapy, combinations of stem
cells, genetically modified cells, modified genes, scaffolding
materials and growth factors have become common tools
●
A major bottleneck in the development of cell therapeutics is the
transition from preclinical research to clinical research on humans
●
Here two different ways of thinking collide:
–
Basic research with large numbers of animals
–
The area of highly regulated clinical studies with humans
●
The question arises how one can prepare researchers for the
requirements of clinical studies in a way that facilitates the transition
of knowledge between basic research and clinical studies

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Introduction
●
All the requirements for clinical trials generally also
apply to clinical trials with biological medicinal products
●
There are also special aspects that must be taken into
account when planning and evaluating clinical studies or
study programs with these drugs
–
Manufacturing, analytical characterization, mechanisms of
drug action and questions of drug safety and toxicology,
questions of environmental safety

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Clinical research vs. Translational research
●
Clinical research
–
Systematic study of human subjects and involving testing of new methods of
diagnosis, prevention and the treatment of diseases
–
Careful and pragmatic testing of mature hypotheses in a controlled environment
●
Translational research
–
Bridging science and practice
–
Linking laboratory science with patients and findings with the needs of the community
–
Possible human applications of scientific and clinical findings
–
Translation into strategies for improving healthcare, patient outcomes, and
community health

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Stakeholders in clinical research
Projects Proof-of-
Concept
Clinical
studies
GMP
production
GMP
production
FinancingFinancing
Industrial
partners
Industrial
partners
Pre-clinical
Research
Academic
partners
Academic
partners
Research
Organisations
Research
Organisations
GCP
oversight
GCP
oversight
Ideas
Theories
Questions
Ideas
Theories
Questions

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Improving pre-clinical trials and translational
research
●
The requirements for successfully conducting clinical trials,
especially for cell therapy studies, are complex
–
Often stem cells, genetically modified cells and nanoparticles have to
be used together
●
The transition from preclinical to clinical testing in humans
represents a bottleneck for academic research
●
We asked ourselves how researchers can be prepared for an
easier entry into the requirements for clinical trials including the
need to work in a regulated area
●
We developed a "reverse engineering" approach that may be
helpful for researchers working at the transition between
translational and clinical research

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The aim of “reverse engineering” approach
●
The “reverse engineering” approach can:
–
Familiarize basic researchers with the requirements and
characteristics of the clinical study culture
–
Prepare the basic researcher to create a more efficient
translational process
●
The goal is preclinical examinations
–
Optimize the regulatory requirements and adaption to the
requirements of GCP
–
Simplify cooperation with clinical study centres, especially with
ECRIN centers and logistics
–
Optimise pre-clinical output for translation in clinical trials

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Basis of the project: Management of industrial-
academical alliances

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The TransVac-Project
●
TransVac Project is a Marie Curie Industry-Academic
Partnership covering:
●
Management of biomedical international research alliances in
Europe
●
Creating knowledge flow through knowledge management
●
Increases the level of research creativity between industrial
and academic biomedical research organizations
●
Creation of a project portfolio to generate knowledge and value
Marie Curie Industry-Academic Partnership
Theme: Trans-national life science value chain management

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Research Alliance of
Transvac

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TransVac-Project: An overview
TransVac was founded in January 2007
●
FP6-MOBILITY - Human resources and Mobility in the specific programme
for research, technological development and demonstration
●
TransVac Management
–
Alliance Office: 4 managers, led by DANDO AND COLUCCI LLC (United
Kingdom)
●
Partners
–
Cambridge University Center for Brain Repair (Cambridge)
–
Ecole Polytechnique Fédérale de Lausanne (Lausanne)
–
ECRIN, Paris
–
Dando & Colucci (Management Company for Innovation Clusters)
–
Fondazione Parco Biomedico San Raffaele (Rome)
–
Coordination by FONDAZIONE PARCO BIOMEDICO SAN RAFFAELE

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Management of the project
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Project management (funds, funding and costs, go no-go decisions, and quality analysis)
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Risk management
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Change and communication management
–
Business managers are "on-site" for the research projects
–
Informal communication between managers based on short term needs
–
Formal communication through web conferences every 2 weeks
–
Monthly executive reports
●
Procedures and hands-on engagement
●
Financial control (collaboration with the administration of the research unit and training on
request)
●
●
●
●
Aim: Reverse engineering of the projects from the intended clinical phase back to the
current state-of-play
Adapted from: J. Dando: International portfolio development and alliances between academia and industry, 2007

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Interactions between
projects

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Management focus on translational research
and interaction with biotechnology SMEs
●
Cooperation with ECRIN working groups in charge of defining tools,
guidelines and procedures to support clinical studies in the EU
●
Special reference to the comparison of national regulatory systems in EU
●
Participation in the development of services supporting phase
1-2 trials on biotechnology-derived products
●
Development of training documents on clinical trial issues in the
biotechnology field
●
Development of a strategy for center selection and fostering patient
recruitment
●
Development of model contracts for EU clinical studies between ECRIN
and biotechnology SMEs, design of a procedure for cost evaluation in EU
clinical studies

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ANALYSIS AND REVERSE ENGINEERING
OF CLINICAL TRIALS REQUIREMENTS
AND PROCESSES

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Method (Information collection)
●
●
Building on the portfolio of the Transvac project, an Europe-wide survey was carried
out within the framework of the ECRIN network
–
to record the requirements and legal regulations for clinical trials with cell products (biotherapy
studies), stem cells, organic products and genetically modified cells
●
Interviews with ECRIN clinical study centers, regulatory authorities, ethics committees
●
Additional surveys were carried out with regulatory authorities
–
Paul-Ehrlich Institute in Germany
–
ISS in Italy
–
AFSSAPS in France
●
Requirements for animal models and for animal experiments
●
Standard Operating Procedures and guidelines of various clinical trials centres were
analysed
●
A concept for the support of preclinical research and a guide for the transition from
preclinical to clinical studies were developed

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Analysis method
Analysis of the use of animal models
–
Animal suppliers, animal husbandry at institutions, non-animal models
●
Analysis and registration of centers that have experience with phase I
studies
●
Collection and analysis of requirements and relevant legal regulations for
clinical trials
–
Focus on clinical trials with cell products, stem cells and genetically modified cells
–
Collecting requirements at meetings, with web conferences and surveys of
researchers and other stakeholders
●
Creation of a concept to support transition from preclinical research to
clinical studies
●
Reconditioning and modification of the analysed material about clinical
trials requirements to fit into the preclinical research environment
●
Development of requirement catalogs, guidelines and training courses

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Reverse engineering
concept

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Specifics of the portfolio projects
●
The portfolio projects include research projects on tissue
regeneration for muscle, heart, bone and nerve cells
●
Genetically modified stem cells are used and partly
supported with the help of biomatrix scaffolds and
growth factors
●
The promotion of neuronal plasticity after spinal cord
injuries is achieved through the use of siRNA and the
introduction of pro-neural genes

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Possibilities for the „reverse engineering“
approach
●
In the complex field of cell and gene therapy, a “reverse
engineering” approach can:
–
Familiarize the basic researcher with requirements and
characteristics of the clinical study culture
–
Considering special conditions for cell and gene therapy studies
–
Development of recommendations for suitable animal models
–
Identify suitable clinical study centers
–
Prepare for working in a regulated environment
●
The goal is preclinical analysis with regard to:
–
Optimizing the regulatory requirements and GCP rules
–
To simplify cooperation with ECRIN study centers and logistics

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Reverse engineering process -
overview
Reverse engineering of the projects from the intended clinical phase
back to the current state-of-play

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TransVac Management
Regularly communication Web conferences
Exchange of information newsletter
TransVac Central Management financing
Managers in the different centres Support of researchers
Exchange between managers
Collaboration with ECRIN Inclusion of EU correspondents
Meetings

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Return of knowledge to basic researchers
Special populations Guidelines and catalogue of
regulations
Recruitment support by ECRIN
Non-animal tests consultation
Good Laboratory Practice (GLP) consultation
Phase I international studies support by ECRIN
Selection of study centers support by ECRIN
Good Manufactoring Practice support by ECRIN
Data protection consultation

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Example: Stem cell clinical
trial requirements

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Example Transvac 08
●
Clinical validation of mesoangioblasts from HLA-matched donors for
treatment of muscular dystrophy
–
Partners: Fondazione Parco Biomedico and ECRIN
–
Phase I clinical study with mesangioblasts from HLA-matched donors
–
Are transplanted into patients who have Duchenne muscle atrophy
–
Treatment was successful in pre-clinical animal models (mouse and dog)
–
Normal stem cells are supposed to fuse with regenerating dystrophin fibers
●
Intra-arterial transplantation tested in phase I study
–
Intra-arterial injections, restoring dystrophin production
Barriers to translational research
See: M Sampaolesi, S Blot, Cossu G, et al. Mesoangioblast stem cells ameliorate muscle function in dystrophic
dogs. Nature 2006; E-pub Nov 15, 2006

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The art of working in a
regulated area
It is especially this requirement that
must be fed back to basic, pre-
clinically working researchers

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Results
●
A lack of understanding was ascertained for the regulatory, ethical and
process management aspects for biotherapy projects
–
Requirements of ethics committees
–
The presentation of own results of research in the context of research state of
the art (i.e. the state of the art)
–
Ability to give a comprehensive and critical overview about research (traceability
by other research groups)
–
The possible toxicity of stem cells in general is underestimated
●
Creation of preclinical evaluation guide for animal models and
requirements for cell and gene therapy products prior to start of clinical
study:
–
Preclinical evaluation, drug safety concerns in phase I studies, EU guidelines for
phase I trials, EMEA guideline for phase I trials for cellular biotherapy and gene
therapy studies, country-specific requirements and regulations at the beginning
of biotherapy studies, overview of the animal models for cell therapy and
regeneration, problem areas of animal studies for clinical trials, animal studies

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Results
●
A process description was developed (stages from preclinical research to phase I
transition process)
–
Analysis of the number of animals and important experimental data in selected cell and
gene therapy studies
–
Legal provisions and advice from the Paul-Ehrlich Institute in Germany and AFSSAPS in
France.
●
List of international requirements for animal models, costs of animals for studies,
special requirements for biotherapy studies, applicable guidelines for tires in
preclinical studies
●
Advice received from competent authorities
–
the national authorities in EU countries and the EMEA recommend attending advisory
meetings prior to the start of phase 1 studies
–
Questions in connection with the new investigational medicinal products, including the
relevance of the animal model
–
But authorities do not make recommendations for a model for any particular disease
–
A similar response in human and animal cells in vitro, does not guarantee that the in vivo
response is similar

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Results of survey of Clinical Trials Centres
Clinical Study Centers (mostly ECRIN Clinical Trials Centres)
–
Little experience in phase I studies
–
Little experience with stem cells
–
No experience with clinical studies of nanotechnology products
–
Little contact and information of preclinical phase
●
Animal breeding and husbandry
–
Only a limited range of animal models at university clinics
–
Little experience with alternative testing methods
–
Use of commercial animal providers
●
Important topics for training preclinical researchers
–
Legal regulations, guidelines, GCP requirements, national differences in
regulation and practice
–
Requirements from authorities and ethics committee, clinical trials process
requirements
–
Data management / data privacy requirements

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Results
●
The right animal model for preclinical research is essential
–
Analysis of the number of animals, characteristics, type of treatments for heart muscle
regeneration, skeletal muscle regeneration, bone development and neuroregeneration
–
Identification of problem areas in animal studies, international requirements for animal models,
costs of animals, measures for animal welfare, non-animal models
●
Collection of requirements for cell and gene therapy products prior to the start of a
clinical study
–
Preclinical evaluation, use of nanoparticles, drug safety concerns in phase I studies,
requirements of the EMEA guideline for phase I studies on cellular biotherapy and gene therapy
and country-specific conditions
●
There are no general rules for considering the number of animals in selected cell and
gene therapy studies
●
There are considerable country-specific differences in the field of:
–
Cell therapeutic requirements for animal models, study implementation, studies with stem cells,
studies with nanoparticles and active ingredients from animal or human products
–
Provision of animal models and the requirements for animal experiments
–
This makes it easier to carry out some aspects of preclinical research or clinical studies in some
countries than in others

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Results
●
Improvements for basic researchers can be achieved
through:
–
Training of the important aspects of good clinical practice,
intensive dialogue with the authorities already during
development phase
–
Careful selection of animal models and non-animal models
–
Inclusion of centers with experience in phase I and / or stem cell
therapy
–
Comprehensive evidence of cellular safety (cell migration, cell
type, pre-neoplastic changes)
–
Complete presentation of published research data
–
Facilitate the conduct of international cell therapy studies in
ECRIN

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Results
●
GCP training covering
–
Phase I-IV studies, differences between commercial and academic studies,
the different categories of clinical studies, the GCP directive, the various
actors (sponsor, study doctor, LKP, EK, authority, etc.), side effects, etc. Study
design, cross-over studies, controlled randomized studies, important terms
such as confounder (disruptive variable) process of informed consent, written
patient information, written patient information, revocation of consent,
declaration on data protection, data protection: personal data, sensitive data ,
transfer of study data, FIM (FIRST-IN-MEN) studies, data and risk
assessment, infrastructure for clinical research: centers and networks, clinical
study centers and clinical study units, clinical research costs
●
Overcome obstacles to the innovation of clinical research
–
Better dialogue with the authorities already during development
–
Increased acceptance of biomarkers and surrogate endpoints by authorities
–
Increased participation of other interest groups such as e.g. patients in the
regulatory review process

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Results
●
Overcome difficulties conducting clinical trials
–
Lack of predictability in the operational environment, qualification
bottleneck
–
Lack of early support
–
Research of degenerative and chronic diseases is in general more
expensive (more complex patient treatment and monitoring is
necessary), long time to observe effects
–
Complex regulatory and ethical requirements
●
Very few centers have experience with phase I and even less
with FIM studies
●
Special features to consider: logistics, the availability of an
emergency rooms, etc.

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Details of the employed Portfolio studies
●
Skeletal muscle
–
Muscle regeneration with either wild type of genetically modified cells
●
Cardiac muscle
–
Cardiac mesoangiobalsts are induced to differ­entiate into bio-matrix scaffolds and
implantted
●
Cardio­vascular
–
Generation of cardiomyocytes from stem cell sources and gene delivery to such
cells
●
Bone formation
–
Skeletal stem cells used in combination with scaffolds and differentiation factors
●
Neurological
–
siRNA approach: reduction of expression of three target genes
–
Delivery pro-neural genes by the use of cationic oligo­peptides and inhibition by
vector-driven siRNA

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Portfolio studies: international requirements for tisssue studies
Requirements for specific populations, blood and tissue samples and genetic
studies by national authorities were collected and analysed. Example:
Country requirements for specific
populations
requirements regarding blood
and tissue samples, circulation
and storage
Specific requirements
regarding genetic
studies
Hungary no Yes, comprehensive listing of
investigations, specific IC for
DNA samples. DNA samples
max storage for n15 years
Yes, specific IC
UK Yes, COREC application form:
www.corec.org.uk
Yes: human tissue act.
www.opsi.gov.uk/acts
/acts2004/20040030.htm
no
France Yes: vulnerable pop (children,
pregnant, dementia,…) Art L
1121-5,-6,-7,-8,-9,-11,-14. L1122-
1,-2
Yes: Art L 1125-1. L 1243-3 Yes: CNIL, Comité
national d’éthique, Loi de
bioéthique
Italy Yes, vulnerable population
(children, unconscious, dementia)
article 3,4,5 legislation decree:
//oss-sper-clin.agenziafarmaco.it
/normative/dlgs_211_24-6-
03_Dir_UE_sper.pdf
yes Yes: specific IC, purpose
must be described in
advance
Spain no Yes: RD 478/1993:
www.agemed.es/actividad
/legislacion/espana/
especiales.htm
no
Ger Yes: vulnerable pop (children,
pregnant, unconcious,…) AMG
§41, MPG §20, §21, §88
StrlSchV, §28 RöV

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Ontologies
Data standards
Biomarkers
Methods
Processes
Analyses
Disease models

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Discussion
●
In the particularly complex area of ​​stem cell-based cell therapy, a
“reverse engineering” approach can facilitate the transition from basic
research and preclinical studies to phase I / II clinical studies
●
By familiarizing basic researchers with the requirements and
peculiarities of the clinical study culture Approval for focused pre-
clinical research allowed
●
Reverse engineering generally refers to the process of extracting the
design elements and processes from an existing system by
examining the structures, states and behaviors
●
We have transfered this concept to clinical studies by examining and
comparing requirements and processes for conducting clinical studies
in 12 European countries in order to develop guidelines and
recommendations for preclinical research in the field of cell therapy

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Discussion
●
Reverse engineering generally refers to the process of extracting elements and
processes from an existing, working system by examining its structures, states and
behavior
●
We transfered this concept to the clinical study area
●
With the new regulation on medicinal products for advanced therapies, the European
Union has created legal certainty in the approval of gene therapy, somatic cell therapy
and tissue engineering products
●
The products for novel therapies (gene therapy, somatic cell therapy and tissue
engineering ) are assigned to drugs area and must therefore pass the same approval
conditions as conventional drugs
●
But SMEs often lack required regulatory know-how for approval and licensing
●
Products from tissue engineering (human tissue engineering medicinal product) are
often developed at universities and research laboratories
●
By 2012 all hTEPs, including those currently in circulation, must undergo approval
procedures by the EMEA
●
Many SMEs fail to meet the requirements of the EMEA in the form of complex clinical
studies as part of the approval process.

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Discussion
●
Suggestions for improvement
●
Training of basic researchers in Good Clinical Practice (GCP)
●
Improved dialogue with the authorities already during the development, e.g. by the
researcher's participation in advisory meetings before the start of the phase 1
studies
●
Verification of relevance of the animal model
●
Checking the migration of cells into the target organs (biodistribution) and cell
differentiation into the desired cell type, but also the absence of preneoplastic
changes
●
Model suitable to demonstrate virus and microbiological safety
●
Support for the comprehensive and complete data collection in support of clinical
studies
●
Consideration of the requirements of the ethics committees and authorities, in
particular with regard to the declaration of consent, data protection (personal data,
sensitive data, transfer of study data abroad) and the special regulations for
participation in clinical studies.

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Contact
Wolfgang Kuchinke
Heinrich-Heine Universität Duesseldorf, Duesseldorf, Germany
wolfgang.kuchinke@uni-duesseldorf.de
wokuchinke@outlook.de
THANK YOU!
This is a translation of the presentation held in German; it contains additional
explanatory material for a workshop.