Can the lung be repaired in emphysema?

Transcription

1 Can the lung be repaired in emphysema? Irina Petrache, MD National Jewish Health and University of Colorado Denver, CO Funding: Disclosures: Scientific founder Allinaire- no conflict with the contents of this presentation Patent applications for ASC use

2 Independently Asthma associated with: Smoking Age Asthma Other risk factors: Genetic predisposition (AATD) Environmental pollution Infections (HIV) Autoimmune disease Spectrum of Obstructive Lung Disease Chronic Bronchitis COPD Emphysema

3 GOLD 2011 COPD severity classification captures disease phenotypes Spirometry (FEV1) Intermediate Mortality Low Mortality High Mortality Intermediate Mortality Frequency of exacerbations Elevated white blood cell count Need for phenotype/endotype specific Pulmonary hypertension approach to COPD Symptoms (quality of life) Augusti A et al. ERJ 2013

4 Ineffective repair/ failure of stem cells Airway and vascular maladaptive Mechanisms of lung injury in COPD Oxidative stress Barrier dysfunction Cell autophagy (mitophagy) Senescence Cell death (apoptosis, necroptosis) Matrix proteolysis Loss of cilia function Abnormal mucus production Inflammation/ Autoimmunity

5 Mechanisms of Repair Interruption of destructive processes Removal of debris: efferocytosis Molecular and cellular repair: autophagy DNA repair plasma membrane repair Restoration of lung maintenance program: Cell regeneration Cell survival Cell proliferation Matrix rejuvenation

6 Respiratory Disease Regenerative Medicine Endogenous Repair: Endogenous Progenitors Exogenous Repair: Gene Therapy Cell Engraftment Functional Repair: Autophagy, Immunomodulation Ex Vivo Tissue/Organ Bioengineering Endogenous lung progenitors ESCs, ips cells MSCs, EPCs Bone marrow mononuclear cells Amniotic fluid cells Emphysema Healthy

7 Exogenous administration of stem/progenitor cells Adult Adipose Stromal Cells (ASC) Angiogenic and anti-apoptotic potential, stabilize blood vessels Secrete GM-CSF, VEGF, HGF, bfgf and TGF-B Rehman J et al, Circulation 04 Traktuev D et al, Circ Res 08 Brief lung homing Anti-apoptotic Cai L et al, Stem Cells 07 Caspase -3 activity (mu/s/ug) Air *p=0.002 **p=0.02 Cig Cig Smk + Smoke ASC Schweitzer et al. AJRCCM 11

8 ASC: protect against CS-induced lung damage CS i.v. ASC Airspace enlargement Schweitzer et al. AJRCCM 11

9 ASC: Paracrine mechanism of action CS ip. ASC

10 ASC protection against systemic CS effects Cachexia Schweitzer et al. AJRCCM 11

11 ASC protection against systemic CS effects BM progenitor cell dysfunction Schweitzer et al. AJRCCM 11

12 Paracrine effects of ASC on BM dysfunction via TSG-6= TNF- stimulated gene 6 Lee, Prokop et al. Cell Stem Cell 09 Lung mrna XIe et al. Stem Cells 2015 CS

13 ASC effect on BM progenitor cells XIe et al. Stem Cells 2015

14 Adult Stem Cells Bone marrow, adipose, cord blood, placenta, etc, Bone marrow stem cells Hematopoietic stem cell (HSC) Endothelial progenitor cells (EPC) Mesenchymal derived stromal cells (MSC) Lanza and Rosenthal Scientific American June 2004

15 CS-induced BM progenitor cell dysfunction in mice Sustained, chronic effects Affects LSK+ stem and progenitor cells 140 BM progenitor cells * * CS (7 wk) i.v. ASC 0 AC CS 4d CS 4d-AC 7d CS (4 d) XIe et al. Stem Cells 2015

16 SDF1-CXCR4 inhibition with AMD3100 ameliorated emphysema in mice Barwinska et al, AJP-Lung in press

17 Mechanisms of Repair Interruption of destructive processes Removal of debris: efferocytosis Molecular and cellular repair: autophagy DNA repair plasma membrane repair Restoration of lung maintenance program: Cell regeneration Cell survival Cell proliferation Matrix rejuvenation

18 Top 5 Dysregulated Pathways Metabolomics+Genomics in COPD Bowler et al. AJRCCM 15 Perturbed Pathway Pathway p- value: Genes Pathway p- value: Metabolites Glycerophospholipid metabolism * 3.70 x x Oxidative phosphorylation * 4.10 x Regulation of autophagy * FCγR-mediated phagocytosis 2.09 x Sphingolipid metabolism 1.40 x x

19 Sphingolipid metabolism

20

21 Human lung microvascular endothelial cells (HLMVEC) Proliferative/progeni tor potential Chronic CS associated with apoptosis-resistant phenotype Petrusca et al AJRCMB 14

22 Chronic CS associated with dysfunctional HLMVEC Pulmonary microvascular endothelial cells (PMVECs) from smokers (S) are less vasculogenic than PMVECs from non-smokers (NS)

23 Exogenous Pro-angiogenic (P) circulating hematopoietic stem/progenitor cells (CHSPCs) may improve HLMVEC function Pro /Non-angiogenic CHSPC ratio reduced in CS associated peripheral vascular disease Estes et al Cytometry 10 Smokers PMVC s ability to form vascular tubes improved by co-culture with pchspcs from non-smokers

24 Pharmacological interventions to improve HLMVEC function

25 Decreased sphingosine kinase activity in COPD lungs Enzyme Activity in Lung Post-BD FEV-1 R (p-value) Severity of Emphysema on CT R (p-value) Sphingosine Kinase 0.28 (0.02) (<0.0001) S1P Lyase 0.26 (0.03) (0.01) S1P Phosphatase 0.26 (0.03)

26 Maceyka & Spiegel Nature, 2014

27 Decreased sphingosine kinase activity in COPD lungs Enzyme Activity in Lung Post-BD FEV-1 R (p-value) Severity of Emphysema on CT R (p-value) Sphingosine Kinase 0.28 (0.02) (<0.0001) S1P Lyase 0.26 (0.03) (0.01) S1P Phosphatase 0.26 (0.03)

28 Decreased S1P1 expression following CS S1P1-dependent effect of S1P agonists (FTY-php) on HLMVEC

29 Effect of increasing S1P signaling in CS-exposed lungs Anti-apoptotic effects of FTY ene-phosphonates CS FTY

30 Decreased airspace enlargement by S1PR1 agonist (GSK)

31 S1PR1 agonists do not restore lung elastance (alveoli), but they improve lung tissue resistance (small airways) Tissue dampening Compliance G(cmH2)/ml) tissue dampening C (ml/cmh2) AC AC CS-c CS-c CSc+FTY-1 CS-s CS-s CSs+FTY-1

32 Effectiveness of S1P Lyase inhibition in mouse lungs

33 Lung inflammation Protective effects of S1P Lyase inhibition on CS-induced mouse lung inflammation and dysfunction

34 Protective effects of S1P Lyase inhibition on CS-induced mouse lung inflammation and dysfunction SPL-inh improves Lung compliance SPL-Inh during CS is not associated with lung fibrosis

35 Improved outcomes of S1P enrichment in CS model of emphysema FTY-phosphonates have distinct or less potent functional effect compared to SPL inhibition Maintenance of intracellular S1P levels may protect lung microvascular endothelial cells from CS-induced injury

36 Proposed Mechanisms of Repair in COPD Phenotype specific Aimed at interrupting self-perpetuating injury Re-engage the lung maintenance program: Cell repair Cell regeneration (resident & recruited progenitor cells) Cell survival

37 John Jung, B.S. Erica Beatman, MS Danting Cao, PhD student Kengo Koike MD, PhD, postdoc Andrew Mikosz Kevin Ni, MD PhD student Kelly Schweitzer, PhD Karina Serban, MD Evgeny Berdyshev, Ph.D. (Head) Irina Bronova, Ph.D. Mass Spectrometry Group at NJH Denver Omics: Russ Bowler Nichole Reisdorph Katerina Kechris IU collaborators: Daniela Petrusca Christopher Poirier (deceased) Merv Yoder Matt Justice Robert Bittman Ed Schuchman Erich Gulbins Rich Kolesnik Tony Futerman Yael Pewzner-Yung

38 Sphingolipids are regulated via metabolism

39 How do organs repair themselves? Endogenous Progenitor Cells Function in development, repair, aging (?) Found in every organ Lung: complex organ >40 cell types Lung progenitors: evolving definitions Most data in mice: less information in humans Homeostasis not well understood Potential role as lung cancer stem cells Many years from clinical application Liu and Engelhardt, PATS 2008 Courtesy of Barry Stripp PhD Cedars Sinai D Kotton Am J. Resp Crit Care Med 2012

40 Dysregulated Airway Progenitor Cells (Basal Cells) play a role in COPD pathogenesis Early events in the pathogenesis of chronic obstructive pulmonary disease: Smoking-induced reprogramming of airway epithelial basal progenitor cells. Shaykiev and Crystal Annals of the ATS 2014

41 Can Isolated Embryonic Stem Cells or Induced Pluripotent (ips) Cells Repair the Lung? Exposure to sequential growth factors in vitro Mimic embryonic development Lung Disease-Specific ips Cells Mou et al. Cell Stem Cell 2012 Somers et al. Stem Cells 2010 No one has yet grown functional lung tissue from ESCs or ips cells

42 Immunomodulation: Mesenchymal Stem (Stromal) Cells Initially isolated from bone marrow Adipose, placenta, cord blood, others Differentiation ability ( stem cell role) Pericytes Line blood vessels, sinusoids Express DAMPs and PAMPs Toll-like receptors Sample and react to inflammatory environments

43 Immunomodulation of Innate and Adaptive Immune Systems By MSCs Immunosuppressive -Inhibit proliferation and effector functions of immune cells in vitro -T cell, B cell, NK cells, dendritic cells, etc Poor Initial Immunogenicity - Low levels of MHC I - No MHC II - No co-stimulatory molecules - Successful use of non HLA-matched allogeneic MSCs Systemic administration - Transiently lodge in lung - Chemotactic/retention at injury sites Nauta, A. J. et al. Blood 2007;110: Clinical Application Clinical trials: graft vs host, Crohn s, MS, diabetes, arthritis cardiac diseases, others Approved in Canada, New Zealand for refractory pediatric GVHD

44 MSC-Based Cell Therapy Approaches for Lung Diseases > 200 publications showing efficacy in pre-clinical models -multiple mechanisms Potential Mechanisms of MSC Actions Cell Contact Independent soluble mediators anti-inflammatory cytokines anti-bacterial peptides others microvesicles inhibitory mirnas Cell Contact Dependent mitochondrial transfer Indirect influence immune cells rather than direct effects on lung cells Weiss DJ, and Rojas M Cell Therapies for Lung Diseases. Stem Cells, 2013