A Physiological Cell based Microchip Platform. 1

Transcription

1 A Physiological Cell based Microchip Platform 1

2 Drug Development Problem What passes here often fails here Discovery Pre clinical Clinical Trials in vitro in vivo Static Well plate Platforms are Not Realistic 2

3 Drug Delivery Process Delivery Vehicle Targeting Ligands Drug Payload Blood Flow Dynamic Process with Active Targeting to: Deliver the Right Drug To the Right Cell In the Right Dose At the Right Time Rolling Receptors Adhesion Release Endothelium Tissue Cells Tumor Cells Drug Delivery Process is Highly Dynamic 3

4 Current Drug Cell Interactions Assays Static Condition Non realistic Scenarios Reagent Hungry Bioreactors 3D Culture Co Culture Lack of in vivo Conditions (Morphological, Physiological) Co culture of Vascular and Tissue Cells Separated by a Diffusion Barrier 4

5 SynVivo Solution Most realistic and dynamic in vitro model More detailed understanding than in vivo Ability to co culture tumor/tissue cells (animal & human) Scalable for high throughput/content Bridging the Gap between Drug Screening and Clinical Trials 5

6 SynVivo Technology Platform Animal/Human Scan Digitized SynVivo Chip SynVivo Assay Simple, Low Cost Fabrication Methods Cell Seeding (~5 min) Cell Culture (12 48 hrs) Experiment (hours-weeks) Simple, Robust Protocols 6

7 SynVivo Applications Basic Research Drug Delivery Drug Discovery ADME / Tox Cultured astrocytes Co-culture of tumor and endothelial cells In Areas of: Oncology Neurology Inflammation White blood cells interacting with endothelial cells Drug particle adhesion Platform Technology for Application specific Assays 7

8 Full Spectrum Shear Adhesion Map from a Single Experiment CAD Model SynVivo Chip Assay Observations Computational Model Shear Rate Map Current practice Large volumes Multiple experiments SynVivo Saves >95% Consumables and >90% Time & Effort Prabhakarpandian B, Pant K, Scott RC, Patillo CB, Irimia D, Kiani MF, Sundaram S, Synthetic microvascular networks for quantitative analysis of particle adhesion, Biomedical Microdevices, 2008, 10 (4),

9 Preferential Adhesion in Bifurcations (in vivo Validation) WBCs binding on inflamed endothelial cells Adhesion Pattern SynVivo Fluorescence image Adhesion Pattern in vivo Fluorescence image SynVivo Captures in vivo Adhesion Patterns Tousi N, Wang B, Pant K, Kiani MF, Prabhakarpandian B, Preferential adhesion of leukocytes near bifurcations is endothelium independent, Microvascular Research, 2010, 80(3),

10 Targeted Drug Delivery (in vivo Validation) Confluent Culture of Endothelial Cells, followed by Activation with TNF α Day 1 Day 3 Day 7 in vivo TNF α activation TNF α activation and particle adhesion in SynVivo Functional Endothelium, Activation, and Targeted Delivery Demonstrated Rosano JM, Tousi N, Scott RC, Krynska B, Rizzo V, Prabhakarpandian B, Pant K, Sundaram S, Kiani MF, A physiologically realistic in vitro model of microvascular networks, Biomedical Microdevices, 2009, 11(5),

11 Screen particles of different shapes Linear Channels Particle Shape Effects (in vivo Validation) SynVivo 240 s 1 in vivo SynVivo Linear channels cannot differentiate between shapes, while SynVivo can N Doshi, B Prabhakarpandian, A Ramsey, K Pant, S Sundaram, S Mitragotri, Flow and Adhesion of Drug Carriers in Blood Vessels Depend on their Shape: A Study using Model Synthetic Microvascular Networks, Journal of Controlled Release, 2010, 146(2),

12 Nanopolymer Tumor Delivery (in vivo Validation) Evaluate two different polymers (A B) complexed with GFP encoding DNA Injection Modes: Direct Injection and Vascular Injection A Direct Injection B Direct Injection A Direct Injection B A B A Vascular Injection B Vascular Injection Vascular Injection Not Possible A B N/A N/A SynVivo Response in vivo Response Well Plate Response SynVivo predicts in vivo response Well plates do not Prabhakarpandian B, Fewell JG, Rea Ramsey A, Pant K, Matar M, Anwer K, Quantifying Gene Delivery Using Synthetic Microvascular Networks. J Control Release. 2015, 201,

13 Blood Brain Barrier (BBB) (in vivo Validation) SynVivo BBB compared against Transwell (current practice) and in vivo Confluent Culture of Endothelial Cells Astrocyte EC Co culture SynVivo BBB SynVivo BBB_ACM SynVivo BBB permeability shows excellent correlation with in vivo permeability! (publication in preparation, data not shown) Permeability Assay Tight Junction and Transporter Proteins SynVivo BBB Exhibits Significant Tight Junction Formation Prabhakarpandian B, Shen M C, Nichols JB, Mills IR, Wegrzynowicz MS, Aschner M, Pant K, SyM BBB: A Microfluidic Blood Brain Barrier Model, Lab on a Chip, 2013, 13,

14 Drug mechanism screening Integrated Inflammation Assay (in vivo Validation) Real time Visualization of Collective or Single Cell Migration Leukocyte Adhesion Cascade in Morphological SynVivo Quantitative Assessment of Drug MOAs Excellent Correlation with in vivo Leukocyte Rolling, Adhesion and Migration in a Single Assay Lamberti G, Prabhakarpandian B, Garson C, Smith A, Pant K, Wang B, Kiani MF. Bioinspired microfluidic assay for in vitro modeling of leukocyte endothelium interactions. Anal Chemistry Aug 19;86(16):

15 Co culture of endothelial and liver cells treated with APAP Organ Specific Drug Toxicity Screening Direct Exposure Vascular Exposure ROS Monitoring Other organs under development Kidney Heart Vascular Lung Nasal epithelium Skin Heart Kidney Lung A Histological Slice Operating in an in vitro Environment Smith A, Chen M, Garson C, Mills I, Prabhakarpandian B, Pant K. Universal Microfluidic Platform for Drug Toxicity Assays (pending publication) 15

16 SynVivo Benefits Realistic Relevant morphology and physiology Co culture capabilities Dynamic Real time Visualization Flow, shear, pressure Save Time/Cost Orders of magnitude reduction in consumables Accelerated development 16

17 SynVivo Availability Products SynVivo Chips Linear Channels Bifurcating Channels Microvascular Networks Co Culture Networks Co Culture Idealized Custom Designs Instrumentation & Accessories Syringe Pumps Microscopes Stage Top Incubator Research Services Bubble free Primer Impedance analyzer Clamps, tubing, etc. Basic research disease mechanisms, biomarker analysis Drug development efficacy, toxicity Drug delivery and screening 17

18 Chip Schematics Vascular Chips Linear Channels Bifurcating Channels Microvascular Networks Custom Designs Co culture Chips Co Culture Networks Co Culture Idealized Custom Designs 18

19 Look Forward to Working Together! 19