Regenerative medicine technologies for therapy and modeling

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1 Institute for Regenerative Medicine Regenerative medicine technologies for therapy and modeling Shay Soker PhD Professor of Regenerative Medicine Wake Forest School of Medicine InterAC Meeting November 24, 2015

2 Research Projects Etc. Cell Therapy (improve self-regeneration) Muscle, Heart Tissue Engineering (replace missing tissues) Cornea, Muscle (+Vascularization; +Imaging), Liver, Pancreas, Etc. Modeling (create human equivalent tissues for research) Liver development and organization, Metabolism and Drug screening, Liver cancer and metastasis Mentoring Funding 2

3 Cell therapy for severe muscle injuries Compartment syndrome (CS) is a severe complication arising from musculoskeletal injuries (fractures, contusions, etc ) CS occurs as the pressure within an osseofascial compartment rises to a level that decreases the perfusion gradient across tissue capillary beds If not treated early, and in severe cases, CS could result in cellular anoxia, muscle ischemia, muscle degeneration, fibrosis and subsequent amputation AFIRM project

4 Rat model of CS; a complex injury Criswell et al AJP

5 Skeletal muscle injury and regeneration (Rats) Remodeling Regeneration Inflammation Degeneration Injury 7D 14D 21D 28D 5

6 Cell therapy: MPC-host interactions Human MPCs into nude rats Baseline muscle function analysis Day 0 CS induction Day 4- MPC injection Harvest tissue (Day 21) 6

7 Cell therapy with human MPCs Human nuclear antigen Ki67 MPCs from older or sick human patients insufficient numbers of stem cells or lack of myogenicity 7

8 Skeletal muscle injury and regeneration in older individuals Inflammation Regeneration Remodeling Degeneration Fibrosis Injury 7D 14D 21D 28D 8

9 The impact of age and gender on MPC therapy; donor-cell:host-environment interactions Young MPCs Adult MPCs Young and adult MPCs Young and adult rats Male and female rats Age matched and mis-matched transplantation Lewis rat MPCs into Lewis rats MPCs labeled with GFP lentivirus Young rats Adult rats Aged rats Baseline muscle function analysis Day 0 CS induction Day 4 - MPC injection Muscle function/ Harvest tissue (Day 14/28) 9

10 Transplanted MPCs Restoration of muscle function Host Tissue Day 14 Peak Isometric Torque/Hz (% Uninjured) PBS AMPCs * YMPCs Young Adult Aged *Functional improvement seen only in adult animals treated with young MPCs. 10

11 Goal: Replace missing and/or damages muscle tissue 11

12 Implantation of bioengineered muscle In vivo imaging Histology Criswell et al Biomaterials

13 Bioreactor Solution: Image regenerating tissue nondestructively in real time in vivo 13

14 Imaging depth is limited > 0.5 mm? MIC ST CM TPM Animal Skin ST: Standard fluo. microscope CM: Confocal microscope TPM: Two-photon microscope Lens && Detector Our solution: Decouple source and detector Embed Micro-Imaging Channel 14

15 Mapping Results Control Image Control Image Mapped Image Mapped Image Hofmann MC et al Tissue Eng Part C 2012, Journal of Biomedical Optics

16 Cornea transplantation Descemet s Stripping and Endothelial Keratoplasty (DSEK) A. B. C. D. A. Corneal endothelial region is damaged (e.g. Fuchs dystrophy) B. Damaged endothelial layer is removed (Descemet s stripping ) C. An endothelial layer with the Descemet s membrane from a donor cornea is removed (a, b) and transplanted into the patient s eye (c). D. The transplanted endothelial layer is fused to the native corneal stroma. 16

17 Development of gel-based scaffolds Gelatin contains multiple glycine (almost 1 in 3 residues, arranged every third residue), proline and 4-hydroxyproline residues. A typical structure is: -Ala-Gly-Pro-Arg-Gly-Glu-4Hyp-Gly-Pro-. Alizarin Red staining (3,000 cells/mm2; 7days after cell seeding) 17

18 Corneal transplantation (Gel scaffolds) 1 day 1 week 2 weeks 3 weeks 4 weeks 5 weeks Na + /K + ZO-1 Normal New Zealand white rabbit (male) Cell seeded scaffold implanted DSEK Observation at 1, 2, 3, 4 and 5 weeks after operation Harvest and evaluation with H&E and IHC Jin-San Choi,.. 18

19 Principles of Tissue Engineering Cell Source Embryonic stem cells Adult stem cells Progenitor cells Signals Growth factors Drugs Mechanical forces Scaffolds Metals Ceramics Synthetic polymers Natural polymers Cells Scaffold Tissue Patient 19

20 Whole Organ Engineering; Limitations Vascularity Organ and tissue complexity 20

21 One approach in whole liver tissue engineering is to use acellular scaffolds RECELLULARIZATION Wake Forest Institute for Regenerative Medicine 21

22 In vivo testing of the whole organ scaffolding system The use of whole organ decellularization for the generation of a vascularized liver organoid. Baptista PM, Siddiqui MM, Lozier G, Rodriguez SR, Atala A, Soker S. Hepatology

23 Potential applications for bioengineered livers Liver biomechanics Liver development Drug testing Disease modeling Liver cancer/metastasis 23

24 Cellular organization; liver regeneration Alb/eNOS/DAPI Mixed Cell Population (HepG2+MS1) Cellular Organization Cell Seeding Day 1 Day 7 HepG2 cells MS1 cells Decell liver parenchyma Decell liver vasculature 24

25 Studying human liver development in vitro using bioengineered liver organoids ALB/CK19 Biliary ducts Hepatocyte clusters 25

26 Bile duct development Human fetal liver Bioengineered liver discs 26

27 Hepatic function 27

28 Body-on-a-chip (Multicellular human tissue constructs) Ex vivo Console of Human Organoids (ECHO) Body-on-a-Chip Drug Screening Platform N C-2027 DTRA ex vivo Capabilities for Evaluation and Licensure (XCEL) program 28

29 There is an urgent need to further understand cancer metastasis 147,000 individuals with invasive CRC each year, with hepatic metastasis occuring in 30% of patients Metastatic growth in distant sights is not well understood Current studies use animal models or insufficient cell culture systems Lack of cell interaction with the tumor microenvironment Joyce, Nature Reviews Cancer, 2009 JohnsHopkins.org 29

30 Colon cancer metastasis is successfully modeled in liver organoid perfusion bioreactor system Day 1 Day 7 Area of Nodules (mm 2 ) 250 ** Day 1 Day 7 Cells in the bioscaffold showed nodule formation around vascular structures (red arrows) Nodules were significantly larger after 7 days 30

31 Modeling of cancer and metastasis R01CA (PQB6) (Agus, Atala, Soker) An Integrative Computational and Bioengineered Tissue Model of Metastasis Wake Forest School of Medicine-Center for Public Health Genomics; Pilot project (Soker) Patient-specific Bioengineered lung tumor organoids (BLTOs) to support personalized medicine. Lance Miller PhD (Cancer Biology, Genomics), Aleksander Skardal PhD (WFIRM, biopronting), Frank Marini (WFIRM, Imaging), Jimmy Ruiz MD (WFU-CCC, oncology), William J Petty (WFU-CCC, oncology), Gregory Kucera PhD (WFU- CCC, tumor bank), Ralph D Agostino (PHS, biostatistics). 31

32 Mentoring Funding Center for Public Health Genomics Tracy Criswell PhD Assistant Professor-WFIRM (Muscle regeneration) Aleksander Skardal PhD Assistant Professor-WFIRM (Tissue biomechanics, Micro-organ models) DoD/The Armed Forces Institute of Regenerative Medicine (Soker) Development of biological approaches to improve the functional recovery after Compartment Syndrome Injury NIH/NHLBI R41 HL (Miller/Soker) ASC101 Enhancement of Regenerative Capacity of Amniotic Fluid-derived Stem Cells Wake Forest University-Translational Science Center (Soker, Marsh, Criswell) Eccentric exercise as a means to improve muscle tissue regeneration post injury in aged rats R01CA (Agus, Atala, Soker) (PQB6) An Integrative Computational and Bioengineered Tissue Model of Metastasis Golfers Against Cancer (Soker/Skardal) A 3-D liver microtumor organoid platform for anticancer drug development Pilot project (Soker) Patient-specific Bioengineered lung tumor organoids (BLTOs) to support personalized medicine N C-2027 DTRA XCEL Ex vivo Console of Human Organoids (ECHO) Body-on-a-Chip Drug Screening Platform Giuseppe Orlando MD PhD Assistant Professor, Surgery-Transplantation (Bioengineered kidney, pancreas, etc.) -Marie Curie FP7-PEOPLE Horizon 2020-BIPCAPAN (2015) NIH/NHLBI R01HL (Soker/Wang) Optical Molecular Tomography for Regenerative Medicine NIH/NIBIB R21EB (Soker,) Nondestructive, High Resolution Imaging Platform for Tissue Regeneration 32