This proposal focuses on the integrative and high throughput functional phenotyping of human blood, matched by Systems Biology and Bioengineering approaches for patient-specific training of computer models to identify and quantify responses to clotting triggers or pharmacological agents. High throughput phenotyping of individual blood samples will be used to train bottom-up and top-down models of blood clotting under static, venous, and arterial hemodynamic conditions.
Specific Aims are:
Aim 1 : Use high throughput intracellular calcium measurements to train neural network models to predict patient-specific response to combinatorial and sequential stimulation, thus testing the milieu that platelets actually experience during thrombosis. Furthermore, high throughput measures of inside-out signaling will be implemented for the development of large scale computational simulation of platelet metabolic pathways.
Aim 2 : Along with platelet phenotyping, we will use validated high throughput blood thrombin phenotyping to identify pathways and synergisms that are defective in patients with existing but undefined defects. These approaches then allow the development of a full platelet-plasma computer simulation of coagulation.
Aim 3 : Using validated tissue factor microarray-flow chambers and microfluidic chambers, we will functionally phenotype thrombus production and clot stability for normal donors and patients under hemodynamic conditions and pharmacological modulation.
Aim 4 : In vivo studies using a mouse laser injury model to follow evolving intrathrombic spatial gradients. The flow studies are supported by advanced multiscale Lattice Kinetic Monte Carlo (LKMC) simulation of clotting under flow using data from all three specific aims. These approaches represent the first full integration of platelet signaling models with realistic and hierarchical hemodynamic/mass transport simulations that regulate adhesive bond function and plasma protease networks. Better elucidation and quantitative measurement of blood reactions and platelet signaling pathways under hemodynamic conditions are directed at clinical needs in thrombosis risk assessment, anti-coagulation therapy during surgery, platelet targeted therapies, and stroke research.

Public Health Relevance

Blood is ideal for Systems Biology research since it is easily obtained from donors or patients, amenable to high throughput liquid handling experiments, and clinically relevant. Clotting and bleeding diseases of aging are seldom due to acquired mutations and this drives the need for advanced functional phenotyping in concert with Systems Biology and other sequencing/genomic approaches.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
5R01HL103419-05
Application #
8688328
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Qasba, Pankaj
Project Start
Project End
Budget Start
Budget End
Support Year
5
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of Pennsylvania
Department
Engineering (All Types)
Type
Biomed Engr/Col Engr/Engr Sta
DUNS #
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Zhu, Shu; Diamond, Scott L (2014) Contact activation of blood coagulation on a defined kaolin/collagen surface in a microfluidic assay. Thromb Res 134:1335-43
Tomaiuolo, Maurizio; Stalker, Timothy J; Welsh, John D et al. (2014) A systems approach to hemostasis: 2. Computational analysis of molecular transport in the thrombus microenvironment. Blood 124:1816-23
Kamat, Viraj; Paluru, Prasuna; Myint, Melissa et al. (2014) MicroRNA screen of human embryonic stem cell differentiation reveals miR-105 as an enhancer of megakaryopoiesis from adult CD34+ cells. Stem Cells 32:1337-46
Li, Ruizhi; Diamond, Scott L (2014) Detection of platelet sensitivity to inhibitors of COX-1, P2Yýýý, and P2Yýýýýýý using a whole blood microfluidic flow assay. Thromb Res 133:203-10
Muthard, Ryan W; Diamond, Scott L (2014) Rapid on-chip recalcification and drug dosing of citrated whole blood using microfluidic buffer sheath flow. Biorheology 51:227-37
Welsh, John D; Stalker, Timothy J; Voronov, Roman et al. (2014) A systems approach to hemostasis: 1. The interdependence of thrombus architecture and agonist movements in the gaps between platelets. Blood 124:1808-15
Stalker, Timothy J; Welsh, John D; Tomaiuolo, Maurizio et al. (2014) A systems approach to hemostasis: 3. Thrombus consolidation regulates intrathrombus solute transport and local thrombin activity. Blood 124:1824-31
Colace, T V; Fogarty, P F; Panckeri, K A et al. (2014) Microfluidic assay of hemophilic blood clotting: distinct deficits in platelet and fibrin deposition at low factor levels. J Thromb Haemost 12:147-58
Dolan, Andrew T; Diamond, Scott L (2014) Systems modeling of Ca(2+) homeostasis and mobilization in platelets mediated by IP3 and store-operated Ca(2+) entry. Biophys J 106:2049-60
Stalker, Timothy J; Welsh, John D; Brass, Lawrence F (2014) Shaping the platelet response to vascular injury. Curr Opin Hematol 21:410-7

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