Theranostic Agents for Diabetes Therapy
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1 Theranostic Agents for Diabetes Therapy Jeff W.M. Bulte, Ph.D. Professor of Radiology, Oncology, Biomedical Engineering, and Chemical & Biomolecular Engineering Director of Cellular Imaging Institute for Cell Engineering The Johns Hopkins University School of Medicine
2 Learning Objectives: 1. Describe the pros and cons of existing non-invasive cellular imaging modalities 2. Understand how microcapsules can provide cellular immunoprotection 3. List 3 imaging modalities that can detect perfluoro-octyl-bromide (PFOB-)labeled capsules 4. Realize the clinical importance of MR-guided injections 5. List at least 2 clinical trials using MR imaging of iron oxide nanoparticle labeled human islet cells OFF-LABEL USE: Feridex (no longer manufactured)
3 Islet Cell Transplantation for Treatment of T1DM
4 Rationale for Encapsulated Cell Therapy Current engraftment rates range from 12-80% Procedure has significant rate of complications Immunosuppressive regimen toxic to islets Microencapsulation may circumvent the need for stringent immunosuppressive regimens
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6 Clinical Imaging Modalities Question Morphology Diagnostics Function Modalities Molecules X-Ray CT MRI Ultrasound Optical Imaging SPECT/PET PET/MRI
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8 Tracking cells after administration: Are they delivered correctly, and effectively? MRI Cell Tracking
9 1. De Vries et al Nature Biotechnology. 23: Zhu et al New England Journal of Medicine. 355;22: Callera et al Stem Cells and Development. 16: Toso et al American Journal of Transplantation. 8: Saudek et al Transplantation. 90;12: Karussis et al Archives of Neurology 67(10): Jozwiak et al. Cell Medicine 2010;1:71 80 As of today there are 7 published clinical MRI cell tracking trials Netherlands 2005 Switzerland 2008 Poland 2010 Czech Republic 2010 Israel 2010 China 2006 Brazil 2007
10 MR Contrast Agent: Superparamagnetic Iron Oxide *
11 MAGNETIC DYE Dextran-Coated Superparamagnetic Iron Oxide Particles (SPIO): Microscopic Magnetic Field Inhomogeneities Induce Rapid Dephasing of Proton Spins and Loss (or Increase) of Imaging Signal B 0 LLS size: ~80 nm
12 MR Tracking of Pancreatic Islet Cells: Direct SPIO-Labeling of Beta Cells 1 wk 18 wks D Jirak, M Hajek et al., MRM 52, (2004) NV Evgenov, A Moore et al., Nat. Med. 122, (2006). Syngeneic Allogeneic J Kriz, F Saudek et al., Transplantation 80, (2005).
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14 Manual counting necessary with high inter-observer variability MRI Results Some patients excluded due to high iron in the liver Low number of hypointense spots compared to amount of islets injected no correlation between #of dark spots and #of islets injected Number of dark signal remained stable over time in all patients
15 Clinical Outcome
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18 Rationale for Encapsulated Cell Therapy Current engraftment rates range from 12-80% Procedure has significant rate of complications Immunosuppressive regimen toxic to islets Microencapsulation circumvents need for immunosuppressive regimen
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20 Capsule Labeling for Immunoprotection and Visualization of Pancreatic Beta Cells Bismuth and Barium Sulfate Superparamagnetic Iron Oxides Perfluorocarbons GadoGold particles CEST agents
21 Use of Semi-Permeable Alginate Capsules for Cellular Immunoprotection
22 Bi X-caps Ba X-caps
23 Feridex-Labeling of APA Capsules: Magnetocapsules No Feridex Feridex Unstained Feridex + Prussian Blue Diameter ~ 350 m
24 Magnetocapsules for Simultaneous Immunoprotection and MR Trackability of Human Islet Cells APA-Feridex APA+Feridex Anti-Dextran/DAPI Human cadaveric islets Murine TC-6 insulinoma DAPI Anti-Dextran Propidium Iodide Newport Green B.P. Barnett et. al., Nat. Med. 13, (2007).
25 MR Imaging Properties of Magnetocapsules Single capsule imaging T2* GRE IRON IRON : positive contrast imaging of capsule surface Imaging of capsule rupture Before After B.P. Barnett et. al., Nat. Med. 13, (2007).
26 MR-Guided Targeted Delivery of MC Human Islets in Swine (Portal Vein Access from Femoral Vein) PV Tip Real-time Pre Immediate Post B.P. Barnett et. al., Nat. Med. 13, (2007).
27 MR-Guided Real-Time Injections Using MR Compatible Injection Catheters Modified loopless antenna design Matching, tuning, and decoupling circuitry Steerable guide catheter via nitinol pull wire 26 gauge nitonol injection needle P. Karmarkar et al., MRM 51, (2004).
28 MR-Guided Targeted Delivery of MC Human Islets in Swine (Portal Vein Access from Femoral Vein) B.P. Barnett, A Arepally et. al., Nat. Med. 13, (2007).
29 3D Imaging of MC Human Islets in Swine Liver B.P. Barnett et. al., Nat. Med. 13, (2007).
30 In Vivo C-peptide Level of Magnetoencapsulated Human Pancreatic Islets Proinsulin = Insulin + C-peptide 2 days post Tx Peri-transplantation phenomena: Islets release insulin storage Human C-peptide level Porcine insulin level Tx=transplantation D.R. Arifin et al.
31 In Vivo Curative Property of Magnetoencapsulated Human Pancreatic Islets in Diabetic Swine Blood glucose O Vetsulin Tx=transplantation Diabetic: Blood glucose > 300 mg/dl Treated diabetic swine did not receive immunosuppressive drugs
32 B.P. Barnett, et al., Radiology 258, (2011). D. Cormode et al., Radiology 258, 1-2 (2011).
33 Perfluorooctyl Bromide PFOB C 8 F 17 Br Known as: Oxygent, LiquiVent MR Contrast Agent Fluorine atoms provide positive contrast under 19 F MRI CT Contrast Agent Residual bromine atom provides significant radioopacity US Contrast Agent 30% acoustic impedence between PFOB and human tissue
34 B.P. Barnett, et al., Radiology 258, (2011).
35 B.P. Barnett, et al., Radiology 258, (2011).
36 B.P. Barnett, et al., Radiology 258, (2011).
37 B.P. Barnett, et al., Radiology 258, (2011).
38 D.R. Arifin et al., Radiology (2011). F.M. Kiessling et al., Radiology 260, (2011).
39 Contrast Agent: (GadoGold) Gd-DTPA is a clinically approved positive contrast MR agent Gold is bioinert and clinically applicable Alric et. al., J. Am. Chem. Soc., 130, (2008)
40 Trimodality of GadoGold Microcapsules Phantoms in Saline Solutions D.R. Arifin et al., Radiology (2011).
41 Trimodality of GadoGold Microcapsules Subcutaneously Injected in Mouse Abdomen GadoGold Saline T 1 -W 9.4 Tesla MRI Micro-CT 40 MHz Ultrasound D.R. Arifin et al., Radiology (2011).
42 In vivo Functionality of Pancreatic Islet Cells Encapsulated in GadoGold Microcapsules Blood Glucose (mg/dl) * Tx Day Treated diabetic mice (n=7) Control diabetic mice (n=8) Healthy mice (n=5) * Statistically different from untreated diabetic mice GadoGold microencapsulated mouse insulinoma βtc6 cells (6000 capsules, 500 cells per capsule) were transplanted into the peritoneal cavity of diabetic D.R. Arifin et al., mice Radiology (2011).
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44 Chemical Exchange Saturation Transfer (CEST) Imaging DIACEST caps: Remote MR Sensing of ph and Cell Viability
45 Chemical Exchange Saturation Transfer (CEST imaging)
46 CEST images and MTR-asym spectra (11.7T) of PS microcapsules at various ph values D.R. Arifin, M.T. McMahon et al.
47 CEST imaging can distinguish live from dead human islet cells Alg/PS/ live cells Alg/PS Alg/PS/ dead cells MTRasym Apoptosis was induced by addition of 10 μm of staurosporin: 30-50% apoptosis ~6% necrosis ph ~ CEST images of PS microcapsules inside 1-mm MR tubes at 11.7 T D.R. Arifin, M.T. McMahon et al.
48 Imaging of Cell Survival using DIACEST capsules KWY Chan, DR Arifin, M.T. McMahon et al.
49 Improving Neovascularization of Islet Transplants: Co-encapsulation of MSCs?
50 XCap Tracking XCaps with MSCs Rabbit Hind Limb Ischemia Model Day 1 Post-occlusion Day 14 Post-occlusion D.A. Kedziorek et al. Stem Cells 30, (2012).
51 XCap with MSCs XCap4 Pre-occlusion Day 14 Post-occlusion D.A. Kedziorek et al. Stem Cells 30, (2012).
52 Enhancing Cell Survival through Co-Encapsulation of (HIF-1a transfected) MSCs and PFOB +MSCs +HIF1-a MSCs +MSCs + PFOB +HIF1-a MSCs + PFOB D.R. Arifin et al.
53 RO1 EB007825
54 MN-siRNA nanoparticles for islet protection iron oxide core (for MRI) sirna Targets: 1. GFP (model target) 2. Caspase-3 3. Beta-2 microglobulin MHC class I light chain sirna Cy5.5 (for near infrared optical imaging)
55 Graft volume MN-siCaspase-3 protects islets from apoptosis Wang P et al, Diabetes, 2011, 60:565
56 Silencing Beta-2 microglobulin (MHC I) with MN-siB2M probe NOD(B2M null)/scid mice MRI, Optical Imaging for 6 wks Day 0 Day 1 Day 3 Day 10 STZ injection (diabetes induction) Islet labeling Islet Transplantation Experimental group: islets labeled with MN-siB2M Control group: islets labeled with MN-control Adoptive Transfer of T cells from NOD mice Western Blot, Histopathology Wang P et al, Diabetes, 2012, in press
57 T2* WI of grafts under left kidney capsule 1 day after AT 1 week after AT 2 wks after AT A. Moore et al.
58 Changes in relative graft volume Delay in diabetes development 17.3 days A. Moore et al.
59 B2M protein expression in the grafts 2 wks after adoptive transfer B2M; Insulin; DAPI A. Moore et al.
60 Learning Objectives: 1. Describe the pros and cons of existing non-invasive cellular imaging modalities 2. Understand how microcapsules can provide cellular immunoprotection 3. List 3 imaging modalities that can detect perfluoro-octyl-bromide (PFOB-)labeled capsules 4. Realize the clinical importance of MR-guided injections 5. List at least 2 clinical trials using MR imaging of iron oxide nanoparticle labeled human islet cells
Magnetic Resonance Imaging of Transplanted Mouse Islets Labeled With Chitosan-Coated Superparamagnetic Iron Oxide Nanoparticles
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