Immunological Challenges of Regenerative Therapies – What ... · Reshaping immune responsiveness...

Immunological Challenges of Regenerative Therapies –What, Why, How ?

Hans-Dieter Volk

Institute of Medical Immunology&

BIH Center for Regenerative Therapies (BCRT)&

Dept. Immunology, Labor Berlin Charité & Vivantes GmbH

Charité – Universitätsmedizin Berlin and Berlin Institute of Health

IMI

Regenerative Therapies are more than Stem Cell therapies

Regenerative Therapies are more than Stem Cell therapies

Regenerative Therapies are more than Stem Cell therapies

Regenerative Therapies are more than Stem Cell therapies

Inflammation/Immunity – a major common challenge

Goal: to support endogenous regeneration and

to improve engraftment and function of biological replacement strategies

Goal: to support endogenous regeneration and

to improve engraftment and function of biological replacement strategies

Challenges: “Aged” immune system, Immunogenicity of Therapeutics, Reshaping Immune Response

Inflammation/Immunity – a major common challenge

Goal: to support endogenous regeneration and

to improve engraftment and function of biological replacement strategies

Challenges: “Aged” immune system, Immunogenicity of Therapeutics, Reshaping Immune Response

Inflammation/Immunity – a major common challenge

Intratissue immunity = part of dynamic tissue homeostasis

immune cells

parenchymalcells

ECM

Tissue

stromal cells

immune cells

parenchymalcells

ECM

Tissue

Challenges (e.g. trauma, infection, (auto)immune attack…)

Recruitmentof cells

stromal cells

Challenged tissue homeostasis triggers recruitment of cells

Immune „Aging“ Naive / early memory T cells Effector T cells

Repetitive antigen(pathogen) challenge

infiltration/inflammation infiltration/inflammation(+) +++

immune cells

parenchymalcells

ECM

Tissue

Challenges

Recruitmentof cells

stromal cells

Enhanced tissue infiltration / inflammation by effector T cells

Immune „Aging“ Naive / early memory T cells Effector T cells

Repetitive antigen(pathogen) challenge

immune cells

parenchymalcells

ECM

Tissue

Challenges

Recruitmentof cells

stromal cells

SPF housing

SPF housing preserves naive immune system despite aging

infiltration/inflammation infiltration/inflammation(+) +++

Immune „Aging“ Naive / early memory T cells Effector T cells

Repetitive antigen(pathogen) challenge

immune cells

parenchymalcells

ECM

Tissue

Challenges

Recruitmentof cells

stromal cells

SPF housingNon-SPF housing

infiltration/inflammation infiltration/inflammation(+) +++

Non-SPF housing brings murine system close to human ones

Age

Frequency of effector T cells in spleen / blood

18 moSPF3 mo

SPF

Non-SPF housing brings murine system close to human ones

Japp A Cytometry 2017, Reinke S Sci Transl Med 2013, Schmidt-Bleek et al. Front Immunol 2017, Sbierski-Kind et al. Front Immunol 2018, Schlundt Front Immunol 2019

Enhanced and decreased level of Teff and Tnaive CD8+ (similar for CD4+, B-Ly, myeloid), respectively, in mice from pet shop (PS) vs. quarantine (Qua) vs. non-SPF vs. SPF housing.

Multiparameter (40) CyTOF analysis

Age

Frequency of effector T cells in spleen / blood

18 moSPF3 mo

SPF

Non-SPF housing brings murine system close to human ones

Japp A Cytometry 2017, Reinke S Sci Transl Med 2013, Schmidt-Bleek et al. Front Immunol 2017, Sbierski-Kind et al. Front Immunol 2018, Schlundt Front Immunol 2019

Enhanced and decreased level of Teff and Tnaive CD8+ (similar for CD4+, B-Ly, myeloid), respectively, in mice from pet shop (PS) vs. quarantine (Qua) vs. non-SPF vs. SPF housing.

Multiparameter (40) CyTOF analysis

Age

Frequency of effector T cells in spleen / blood

18 moSPF3 mo

SPF

3 monon-SPF

12 monon-SPF

Non-SPF housing brings murine system close to human ones

Japp A Cytometry 2017, Reinke S Sci Transl Med 2013, Schmidt-Bleek et al. Front Immunol 2017, Sbierski-Kind et al. Front Immunol 2018, Schlundt Front Immunol 2019

18 monon-SPF

Tnaive and Tmemory/effector cell counts

Japp A Cytometry 2017, Reinke S Sci Transl Med 2013, Schmidt-Bleek et al. Front Immunol 2017, Sbierski-Kind et al. Front Immunol 2018, Schlundt Front Immunol 2019

Fast Test of ImmunoAge - Flowcytometry

Age

Frequency of effector T cells in spleen / blood

18 moSPF3 mo

SPF

3 monon-SPF

12 monon-SPF

Non-SPF housing brings murine system close to human ones

20-50 µl

Non-SPF

SPF

18 monon-SPF

Age

Frequency of effector T cells in spleen / blood

18 moSPF3 mo

SPF

3 monon-SPF

12 monon-SPF

ImmunoAging has a strong impact on thecourse of distinct disease models:

- Organ Transplantation- Spontaneous abortions

- Bone fracture healing- Acute muscle injury- Type 2 Diabetes- Acute Ischemia/Reperfusion Injury- …

- Gene therapy

Non-SPF housing brings murine system close to human ones

Japp A Cytometry 2017, Reinke S Sci Transl Med 2013, Schmidt-Bleek et al. Front Immunol 2017, Sbierski-Kind et al. Front Immunol 2018, Schlundt Front Immunol 2019

Katharina Schmidt-Bleek

Simon Reinke

Christian Bucher

Julia Sbierski-Kind

Sven Geissler

Andreas Thiel

Advanced Preclinical in vitro & in vivo Models

Advanced ModelsClinically relevant

models

Large aimals

Humanized NSG mice& ImmunoAged mice

Biosamples fromdisease-specificpatient population

Multiorgan 3D-chips„Patient-on-the-Chip“

Limited predictive valuefor diseases

Higher predictive valuefor diseases

Conventional preclinical modelsusing mice kept under

SPF housing conditions and biosamples from healthy donors

Goal: to support endogenous regeneration and

to improve engraftment and function of biological replacement strategies

Challenges: “Aged” immune system, Immunogenicity of Therapeutics, Reshaping Immune Response

Inflammation/Immunity – a major common challenge

ImmunoTherapies – innovation drivers in medicine

Restorating ProtectiveImmune Response

Fighting cancer and severe infections

Reshaping UndesiredImmune Response

PreventingSelf-destruction or

Transplant rejection

„Living Drugs“ (ATMP)

Biologicse.g. checkpointinhibitors

e.g. anti-IL-17 mAb

e.g. CAR-T cells

e.g. Treg cells

ImmunoTherapies – innovation drivers in medicine

Restorating ProtectiveImmune Response

Fighting cancer and severe infections

Reshaping UndesiredImmune Response

PreventingSelf-destruction or

Transplant rejection

„Living Drugs“ (ATMP)

Biologicse.g. checkpointinhibitors

e.g. anti-IL-17 mAb

e.g. CAR-T cells

e.g. Treg cells

ReSHAPING undesired immune reactions by regulatory T cells (Treg)

Current strategy:Control of dominant immunopathology bychronic multi-drug treatment

1DG INTERNAL POLICIESWorkshop 2017

Autoimmune Diseases

Solution: Reshape immune balance by Regulatory T cells (Treg)

Treg cells

One Master Cell Product –Many Applications

Aim:ReSHAPEimmune balance

Problem:Increasing prevalence of immune diseases (>10% of chronic diseases), Burden: >100 bn €/a EU1

Current strategy:Control of dominant immunopathology bychronic multi-drug treatment

1DG INTERNAL POLICIESWorkshop 2017

Autoimmune Diseases

Solution: Reshape immune balance by Regulatory T cells (Treg)

Treg cells

One Master Cell Product –Many Applications

Aim:ReSHAPEimmune balance

Problem:Increasing prevalence of immune diseases (>10% of chronic diseases), Burden: >100 bn €/a EU1

ReSHAPING undesired immune reactions by regulatory T cells (Treg)

ReSHAPING undesired immune reactions by regulatory T cells (Treg)

PetraReinkePI OneStudyONE nTreg13

Exemplary Case: My life is better by reshaping immune system

1995 2015 03 / 2015 04/2015 10/2015 2019

ChronicKidneyFailure Dialysis

T1 Diabetes

KidneyTransplan-

tation

Standard of Care

Lifelong Multidrug Therapy(>30 pills/day)

adverse effects, costs, quality of life

Cardio-myopathy

1995 2015 03 / 2015 04/2015 10/2015 2019

ChronicKidneyFailure Dialysis

T1 Diabetes

KidneyTransplan-

tation

Lifelong Multidrug Therapy(>30 pills/day)

adverse effects, costs, quality of life

Cardio-myopathy

RegulatoryT cell

Therapy Minimizing Immunosuppression⇒ 3 pills/d instead >30 pills/d⇒ safe (no infection, no cancer)⇒ excellent graft function (creatinine 1.1)

x Standard of Care

Exemplary Case: My life is better by reshaping immune system

Promising clinical results in LD kidney transplant patients⇒ Safe⇒ hints of efficacy

triple immunosuppression=> FK monotherapy

W1 W24 W60 Y3

Switch to FK monotherapy in the Treg therapy group is feasible(FIH phase I/IIa study)

1st generation of polyclonal Treg – safe and hints of efficacy

Refined Translation – from bed to bench and back to bed next generation regulatory T cell (Treg) approaches

Next-GenerationTreg

Epigenetic and gene editing(CRISPR/Cas)

Gene modified (lentiviral)

Refined Translation – from bed to bench and back to bed next generation regulatory T cell (Treg) approaches

H2020

Next generation T cell products:(M. Schmück-Henneresse / D. Wagner / L. Amini)

Next-generation CRISPR/Cas modified CNI - resistant Treg

Problem: Basal immunosuppression

(tacrolimus) inhibits also Treg

Next generation T cell products:(M. Schmück-Henneresse / D. Wagner / L. Amini)

Effective in vitro expansion and CTLA4 Expressionof KO-TREG under Tacrolimus but inhibition by CSA (safety switch)

Next-generation CRISPR/Cas modified CNI - resistant Treg

Vector-freeCRISPR/Cas9-mediated

FKBP12-KO

d7 of TREG culture

1 2 3 4 5

Tacrolimus-resistant FKBP12–/– TREG

FoxP3

CD25

11.2% 99.1%

High purity sort

Problem: Basal immunosuppression

(tacrolimus) inhibits also Treg

Gundry MC et al.Cell Report 2016

Next generation T cell products:(M. Schmück-Henneresse / D. Wagner / L. Amini)

Effective in vitro expansion and CTLA4 Expressionof KO-TREG under Tacrolimus but inhibition by CSA (safety switch)

Next-generation CRISPR/Cas modified CNI - resistant Treg

Vector-freeCRISPR/Cas9-mediated

FKBP12-KO

d7 of TREG culture

1 2 3 4 5

Tacrolimus-resistant FKBP12–/– TREG

Problem: Basal immunosuppression

(tacrolimus) inhibits also Treg

FoxP3

CD25

11.2% 99.1%

High purity sort

Gundry MC et al.Cell Report 2016

ProTacFIH-Trial2020/1

Goal: to support endogenous regeneration and

to improve engraftment and function of biological replacement strategies

Challenges: “Aged” immune system, Immunogenicity of Therapeutics, Reshaping Immune Response

Inflammation/Immunity – a major common challenge

Problem: Pre-immunity to SpCas9

SpCas9

MHC

TEFF

SpCas9 fragments

Gene-edited CellSpCas9-peptide presentation

Pre-formed SpCas9-reactive Teff cellsregonize CRISPR/Cas edited cell products

⇒ Release of inflammatory cytokines⇒ Killing of gene-edited cells

Problem: Pre-immunity to SpCas9

SpCas9

MHC

TEFF

SpCas9 fragments

Gene-edited CellSpCas9-peptide presentation

Pre-formed SpCas9-reactive Teff cellsregonize CRISPR/Cas edited cell products

⇒ Release of inflammatory cytokines⇒ Killing of gene-edited cells

Test for detecting„fading-out“ Cas peptides

Cas-specifc Treg

Problem solutions

Problem: Pre-immunity to SpCas9

SpCas9

MHC

TEFF

SpCas9-reactive regulatory T cell

TREG

SUPPRESSION

SpCas9 fragments

Gene-edited CellSpCas9-peptide presentation

SpCas9-specific Treg cellsregonize CRISPR/Cas editedcell products andinhibit

⇒ release of inflammatorycytokines

⇒ killing of gene-edited cells

Conclusions for BIH/Charité Translation Strategy

1. SPF housing limits the value of many experimental models as predictive disease models⇒Need for controlled „dirty“ housing at Charité/BIH

2. Reshaping immune responsiveness is a key element to support endogenous regeneration as well asengraftment of tissue replacement approaches that requires in-depth immune biomarker analyses

⇒Need for high-end immune biomarker unit (spin-off CheckImmune)

3. Immune (cell) therapy is one of the most dynamic fields in medicine with high translational potencyat academic centers

⇒Need for developing a Hub (BeCAT + spin-off incubator + biotech/pharma) on AdvancedTherapies at Charité/BIH

Defined, chemically produced Small Molecules

since >120 years

Protein-based drugsproduced in living cells

Biologics

since >30 years since >10 years

chance of cure(game changer)

complexity

“Living” drugs(somatic cells, gene modified cells, in vivo gene therapy, engineered tissues)

Advanced Therapies (ATMP)

Complexity of chronic diseases requires complex therapeutic approaches

The Novel Class of „Living“ Drugs (Advanced Therapies) – a Game Changer

1,069Clinical Trials

underway worldwideby end of Q2 2019

Phase I: 358 Phase II: 617 Phase III: 94

Gene Therapy Gene–modified Somatic Cell TissueCell Therapy Therapy Engineering

366 410 249 44

Alliance Regenerative Medicine Report Q2 2019

Advanced Therapies – not just a dream, it is already “reality“

2019: about a dozen products approved in Europe (marketing authorization)

drug discovery

business models

supply chain

clinical trialdesign

health economy/reimbursement

drugs raw material

scaling-up

pharmacokineticspharmacodynamics

manufacturingmode-of-actionmultiple effects

ethics

complex dataat all TRLs

preclinicalmodels

„Living“ Drugs (AT) – a disruptive innovation shattering current paradigms

§ 91bWissenschaftsrat 2017

Concept ranked as 1st

30 Mio € for the „hardware“GMP-Research Building

Late Clinical Research and Marketing Authorization

Industry PartnershipPharma & Biotech

& Technology Providers

Spin-off of BCRT: Berlin Center for Advanced Therapies (BeCAT)

Prof. Dr.med. Petra Reinke

MoreFasterSuccessfulAffordableAccessible

High need for targeted efforts and a sustainable large-scale research initiative in Europe

Mission:Building-up an ecosystem to

Overcome the technological and regulatory roadblocks for Advanced Therapies in Europe andDeliver a pipeline of dozens of revolutionary Advanced Therapy products by 2030

outgrowingcommercial infancy

Who is behind the RESTORE community ?

Key Elements to support Advanced Therapies in a new way

Infrastructure(Translational Hub´s)

Structured Research and Innovation Actions (RIAs)

Technology Research& Innovation Platforms

Private Public Partnershipand innovative SMEs

Coordination & Support Action(CSA)

Suitable Horizon Europe Programme Elements for RESTORE:- European Regional Development Fund- European Innovation Ecosystems & EIT Health- European Innovation Council (EIC)- RIA´s within Cluster Health- Partnership Innovative Health Initiative- Mission Cancer

Academic, Industry, Patient and Public Society

transparent and dynamic governance

Bilateral interaction with

Regulatory Bodiesat national/EU level

https://restore-horizon.eu

Conclusions for BIH/Charité Translation Strategy

1. SPF housing limits the value of many experimental models as predictive disease models⇒Need for controlled „dirty“ housing at Charité/BIH

2. Reshaping immune responsiveness is a key element to support endogenous regeneration as well asengraftment of tissue replacement approaches that requires in-depth immune biomarker analyses

⇒Need for high-end immune biomarker unit (spin-off CheckImmune)

3. Immune (cell) therapy is one of the most dynamic fields in medicine with high translational potencyat academic centers

⇒Need for developing a Hub (BeCAT + spin-off incubator + biotech/pharma) on AdvancedTherapies at Charité/BIH

4. Immunology is a major common challenge for Regenerative Therapies and a research focus at theBCRT with internationally recognized USP

⇒Need for adequate appointment policy

GMPPetra ReinkeSybill Landwehr KenzelDaniel KaiserAndy RoemhildCarola Beier Henrike Führer Anne ForkeInsa Lehmann …

Biomarker & PreclinicsBirgit SawitzkiNina BabelMichael Schmueck-HenneresseDimitrios WagnerLeila Amini, Ghazaleh ZarrinradDesiree Jaqueline WenderingMathias Streitz, Kerstin Jülke,Gerald Grütz, Levent AkyüzChristian Meisel + LB Immunology Team

Clinical Trial(s)Petra ReinkeMohamed Abou El-EneinSybill Landwehr-KenzelAnett SefrinCordula Giesler…

Physicians & Nurses Patients

Funding (big elephant in the room)Federal Ministry for Education & Research (BMBF) European Union FP7 and H2020 projectsGerman Council of Science and Humanities §91b

Acknowledgement

Patients