MSCs for ARDS Promise and Pitfalls

Transcription

1 MSCs for ARDS Promise and Pitfalls John Laffey Department of Anesthesia, St Michael s Hospital, University of Toronto, CANADA

2 Disclosures Funding European Research Council [FP-7] Health Research Board [Ireland]

3 Bone Marrow derived Stem Cells

4 Nemeth et al, Nature 2009; 15: 42-9

5 C57BL/6 Mice Randomised CLP Sepsis CLP Sepsis 1 million Allogeneic BMSC s Control Allogeneic cells Injury Severity Survival Injury Severity Survival Nemeth et al, Nature 2009; 15: 42-9

6 MSC s increase survival Nemeth et al, Nature 2009; 15: 42-9

7 Gupta, Krasnodembskaya et al, Thorax 2012

8 8 h BAL E. Coli cfu (X10 3 /ml) 24 h Lung E. coli cfu (x 10 5 /ml) Antibacterial action of MSCs A B *, # * MSC 3T3 PBS 0 PBS MSC Gupta, Krasnodembskaya et al, Thorax 2012

9 Chimenti et al, eur Resp J 2012

10 Cell based Gene Therapy Using Cells to deliver genes to lung Using Genes to target Cells to lung Using Genes to modulate cell differentiation Combining Genes and Cells for therapeutic effect

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12 Don t worry. I had the same thing and they cured me!

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14 Lee et al, PNAS 2009

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16 Stem Cell Therapies focusing on lung Repair?? Ware L, Matthay M, NEJM 2000

17 Curley et al, Thorax 2012 Experimental Plan Anaesthetise Intubate 2 million MSCs 2 million MSCs Anaesthetise 24hrs 24hrs Baseline Ventilation 20 min VILI 35cmH 2 O Compliance ed 50% Recover and extubate Surgery, Tracheostomy, Assessment of ALI

18 Curley et al, Thorax 2012 MSCs enhance Repair following VILI

19 Curley et al, Thorax 2012 MSCs modulate inflammatory response following VILI

20 Curley et al, Thorax 2012 MSCs enhance injury resolution following VILI

21 Sprague Dawley Rats Randomized VILI VILI VILI VILI Vehicle 2 million fibroblasts RECOVERED AND EXTUBATED 2 million MSCs Conditioned medium Vehicle 2 million fibroblasts 24 HOURS 2 million MSCs Conditioned medium Degree of Recovery 48 HOURS Degree of Recovery

22 How did the MSCs work? Cells Secretome Curley et al, Thorax 2012

23 Curley et al, Unpublished Data Intravenous vs Intra-tracheal MSC therapy

24 Curley et al, Unpublished Data Intravenous vs Intra-tracheal MSC therapy

25 Alveolar Cellular Infiltrate Curley et al, Unpublished

26 Human MSCs in VILI Repair

27 Sprague Dawley Rats Randomized VILI VILI VILI VILI Vehicle 4 million fibroblasts RECOVERED AND EXTUBATED 4 million hmscs 4 million hmscs 24 HOURS Degree of Recovery Degree of Recovery

28 hmscs enhance Lung Repair Hayes et al, unpublished Data

29 Lowest Effective hmsc Dose Hayes et al, unpublished Data

30 How do MSCs work?

31 Kotton D et al. Development 2001;128:5181

32 Nemeth et al, Nature 2009; 15: 42-9

33 MSCs reprogram Macrophages Nemeth et al, Nature 2009; 15: 42-9

34 Krasnodembskaya et al, Stem Cells 2010

35 MSC anti-microbial effect LL-37 Dependent LL-37 Dose Krasnodembskaya et al, Stem Cells 2010

36 MSCs also make Lipocalin 2 Gupta, Krasnodembskaya et al, Thorax 2012

37 Epithelial Wound Repair Standard medium Fibroblast conditioned medium Alveolar epithelial cells (A549s) MSC conditioned medium MSC co-culture

38 Role of KGF Curley et al, Thorax 2012

39 Islam et al, Nature 2012

40 Islam et al, Nature 2012

41 MSCs for ARDS - Potential Low immunogenicity potential allogeneic USe Can evade clearance by host immune system Some evidence for host response to MScs Potent immune modulatory effects Multiple immune active agents secreted [IL-ra; IL-10; TSG-6; PGE2 etc] Secrete variety of growth Factors KGF; HGF; DGF etc Potential to reduce inflammation and enhance repair processes Efficacy in multiple relevant models [mmsc; rmsc; hmsc] Systemic Sepsis Pneumonia VILI IR

42 Pitfalls what is the key Mechanism? Role of Paracrine Mechanisms Elements of MSC secretome KGF Anti-microbial Peptides Prostaglandin E2 [? + Others] Immune system reprogramming Role of Contact dependent versus independent mechanisms MSC RNA/Mitochondrial Transfer Transdifferentiation [very unlikely] Multiple competing Mechanisms of Action exist Is the MSC Mechanism of action dependent on Injury Type? Are these mechanisms complementary or competing? How important is the micro-environment to mechanism of action?

43 Other key Translational Gaps Should we give MSCs or just give the key Factors e.g. KGF? What is the optimal delivery route(s)? Intravenous versus Intra-tracheal versus Other Does this vary depending on e.g. etiology of Injury How well do MSCs work in more complex preclinical models? What is the optimal dosage regimen and therapeutic window? Address concerns in relation to potential downsides Fibrosis? MSC fate in humans? Ware and Matthay, NEJM 2000

44 Stem Cells for ARDS Progress Report ARDS good candidate disease Need to learn much more about biology of ALI Enable harnessing of potential of these approaches Stem cell therapies offer considerable potential for ALI Pre-clinical studies highly encouraging Closer to clinical testing Danger of attempting clinical translation in advance of better understanding of mechanisms of action Ware and Matthay, NEJM 2000

45 Acknowledgements Research Fellows Donall O Croinin Alistair Nichol Martina Ni Chonghaile Brendan Higgins Joseph Costello Patrick Hassett Daniel O Toole Claire Masterson Maya Contreras Patricia McHale Bilal Ansari Gerard Curley Mairead Hayes Funding European Research Council [Framework 7 Programme] Health Research Board [Ireland] Yamanouchi European Research Foundation Association of Anaesthetists of Great Britain and Ireland St Michael s Hospital Research Foundation