Nanoparticle-based fluorescence molecular imaging Farouc Jaffer MD - PowerPoint PPT Presentation

Nanoparticle-based fluorescence molecular imaging Farouc Jaffer MD PhD Cardiovascular Research Center, Cardiology Division Massachusetts General Hospital, Harvard Medical School Friday April 19, 2018. 08:40 – 10:10 SESSION: INNOVATIONS FOR PLAQUE IMAGING

Basis for Coronary Molecular Imaging • Coronary plaque and stent complications remain leading causes of morbidity and mortality worldwide • Structural imaging of high-risk plaques (e.g. IVUS) currently do not predict risk well enough to enable clinical action. The PROSPECT IVUS-VH trial showed a PPV of only 18% to predict ACS at 3 yrs. 1 • Molecular imaging of coronary plaque biology offers a new approach to understand drivers of plaque and stent complications 2 • Noninvasive coronary molecular imaging approaches are limited by resolution and motion 1 Stone GW et al. NEJM 2011;364:226-35 2 Mulder WJ, Jaffer FA, Fayad ZA, Nahrendorf M. Sci Transl Med 2014

Intravascular Near-Infrared Fluorescence (NIRF) Imaging: A High-Resolution Pathway To The Coronary Arteries • Optical imaging is in the Cath Lab : Optical coherence tomography, near-infrared spectroscopy, angioscopy • NIR window for fluorescence : ↓photon --Weissleder, Ntziachristos Nat Med 2003 attenuation (more light penetration) ↓autofluorescence (less background) • NIRF Molecular Agents : Indocyanine green, Prosense VM110, Bevacizumab-IR800; NIR Fluorecscent nanoparticles? --Kim, Yoo.. Jaffer, T earney, Nat Med 2011 - Osborn et al. JACC CV imaging 2013;6:1327. Jaffer, Verjans. Heart 2014;100:1469; Bourantas et al. European Heart Journal 2017

Nanoparticles (NPs) as molecular imaging platforms Attributes • Scale (1-100nm) facilitates NP interaction with cells • Controllable physical properties and surface modifications for ligand attachment, and longer circulation time • Multimodal imaging capable • Integrated therapeutic-diagnostics (“theranostics”) Limitations • Few are clinically approved • Longer circulation time limits point-of-care applications

USPIO: Clinically Tested Nanomaterials for Imaging of Plaque Inflammatory Cells (Macrophages) • USPIO: Ultrasmall super-paramagnetic PRE POST iron oxide (~30 nm) • Targets plaque macrophages • Strong R2>R1 effects ICA • Clinically tested N>300 pts; Long- circulating; Peak CNR 36 hrs Iron Mac --Kooi et al. Circulation 2003; Trivedi et al. Stroke 2004; Neurology 2006; Stroke 2007

VCAM-1 dual-modal MRI-optical STATIN HCD nanoparticles to assess anti- 240 inflammatory atheroma therapy MRI 160 • VCAM-1 is an important inflammation target * Optical •Phage-display derived VCAM-1 targeting peptide post VCAM-1 •USPIO scaffold derivatized with 8- 12 VCAM-1 targeting peptides 5 * • Tested in ApoE -/- ± statin --Nahrendorf, Jaffer, Kelly..Weissleder Circulation 2006

Open questions: What atheroma cell populations are actually targeted by USPIO NPs? What mechanisms govern NP targeting in vivo? • While most atheroma studies suggest Iron Oxide NPs (USPIO) target macrophage, a number of studies suggest that non- macrophage atheroma cells are also available targets for USPIOs. CLIO-CyAm7 • As FDA-approved USPIOs such as Feraheme are not fluorescent, we prepared a NIRF USPIO (CLIO-CyAm7) for investigation in vivo in atheroma-bearing rabbits susceptible to thrombosis -- Jason McCarthy, MGH Center for Systems Biology

NIRF iron oxide NP targeting of atheroma • N=31 NZW rabbits with A aortic atheroma CLIO WL • IV CLIO-CyAm7 2.5-5.0 Fe/kg 24 hours before NIRF imaging B • Macroscopic PBS WL fluorescence reflectance imaging and fluorescence NIRF microscopy demonstrates C D E F C uptake in atheroma CLIO PBS -- Stein-Merlob AF, Hara et al. Circulation: Cardiovascular Imaging 2017

CLIO NP targets multiple plaque cell types; at the surface and in deeper areas of neovessels B1 A1 CLIO-CyAm7 CLIO-CyAm7 Intimal- Superficial Medial Intima Border (2/3) EC (1/3) L M C D L CD31 CD31 3 1 CD31 B2 A2 Mac L L L RAM11 RAM11 R A M 1 1 RAM11 B3 A3 SMC L a L aSMA aSMA S M A aSMA -- Stein-Merlob AF, Hara et al. Circulation: Cardiovascular Imaging 2017

CLIO NP deposition occurs in areas of plaque permeability A B H&E EB CLIO • Subset of NZW rabbits AF • IV CLIO-CyAm7 5.0 Fe/kg, 24 hours prior • Co-injected with Evans Blue C D permeability marker, 6 mL 0.5%, 30 minutes prior to sacrifice • Strong relationship between depth of penetration between CLIO E F and Evans Blue (r=0.64)

Does CLIO NP deposition identify plaques susceptible to thrombosis? Pre-trigger • NZW rabbits with atheroma • IV CLIO-CyAm7 2.5 Fe/kg, 24 hours prior Post-trigger • Intravascular 2D NIRF molecular imaging and IVUS Thrombus • Constantides model of triggered thrombosis over 48h (RVV, Ex Vivo histamine) • Follow-up intravascular 2D NIRF H * and IVUS at 48h -- Stein-Merlob AF, Hara et al. Circ: Cardiovascular Imaging 2017

CLIO-CyAm7 deposition is greater in plaques that develop thrombosis •Triggered- C A Pre Post D B Post thrombosis • IVUS and Thrombus Carstairs verified thrombosis in 67% Carstairs’ • Detailed E F G H fluorescence microscopy * • Greater NP in plaques with thrombosis CLIO RAM11 AF -- Stein-Merlob AF, Hara et al. Circulation: Cardiovascular Imaging 2017

Conclusions • Molecular imaging of NIR fluorescent iron oxide NPs reveal: - NP deposition is modulated by plaque permeability - NPs target macrophages, smooth muscle cells and endothelial cells within atheroma - NPs are more intensely localized in plaques that are susceptible to triggered thrombosis - NPs can be detected by intravascular NIRF catheters, offering a pathway towards coronary artery detection of plaques predisposed to thrombosis.

Acknowledgements MGH CVRC/Cardiology Technical University of Munich MGH Wellman Center for Ion Botnaru Dmitry Bozhko Photomedicine Marcie Calfon Vasilis Ntziachristos Hongki Yoo Jie Cui (ASN) Giovanni Ughi MIT Amr El Maghraby Joe Gardecki Aditya Kalluri Tetsuya Hara (AHA) Zhonglie Piao Elazer Edelman Stephan Kellnberger (AHA) Kanwarpal Singh MGH-CSB Chase Kessinger (T32) Gary Tearney Jason McCarthy Jin Won Kim, Prof. BWH Cardiology Charles Lin Adam Mauskapf Elena Aikawa Yoshi Iwamoto Eric Osborn (T32, KL2, K08) Masanori Aikawa Claudio Vinegoni Ashley Stein-Merlob Eduardo Folco Ralph Weissleder (HST 15, AHA) Hiroshi Iwata Johan Verjans (Rubicon) Amelie Vromann Funding Ahmed Tawakol Peter Stone -NIH R01 HL108229, R01 HL122388 MGH Vascular Surgery -AHA Grant In Aid, Peter Libby Richard Cambria -HHMI Career Development Award -Canon, Siemens, Merck, Kowa Univ. of Michigan -MGH CIMIT, SPARK, ECOR, CVRC grants Peter Henke -MGH Hassenfeld Scholar Award

Thank you for your attention Farouc Jaffer MD PhD fjaffer@mgh.harvard.edu; @faroucjaffer