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)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL103419-03
Application #
8293284
Study Section
Special Emphasis Panel (ZRG1-VH-D (50))
Program Officer
Qasba, Pankaj
Project Start
2010-09-01
Project End
2015-06-30
Budget Start
2012-07-01
Budget End
2013-06-30
Support Year
3
Fiscal Year
2012
Total Cost
$740,418
Indirect Cost
$242,104
Name
University of Pennsylvania
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Zhu, Shu; Lu, Yichen; Sinno, Talid et al. (2016) Dynamics of Thrombin Generation and Flux from Clots during Whole Human Blood Flow over Collagen/Tissue Factor Surfaces. J Biol Chem 291:23027-23035
Zhu, Shu; Tomaiuolo, Maurizio; Diamond, Scott L (2016) Minimum wound size for clotting: flowing blood coagulates on a single collagen fiber presenting tissue factor and von Willebrand factor. Integr Biol (Camb) 8:813-20
Li, Ruizhi; Elmongy, Hanna; Sims, Carrie et al. (2016) Ex vivo recapitulation of trauma-induced coagulopathy and preliminary assessment of trauma patient platelet function under flow using microfluidic technology. J Trauma Acute Care Surg 80:440-9
Brass, L F; Diamond, S L (2016) Transport physics and biorheology in the setting of hemostasis and thrombosis. J Thromb Haemost 14:906-17
Zhu, S; Welsh, J D; Brass, L F et al. (2016) Platelet-targeting thiol reduction sensor detects thiol isomerase activity on activated platelets in mouse and human blood under flow. J Thromb Haemost 14:1070-81
Diamond, Scott L (2016) Systems Analysis of Thrombus Formation. Circ Res 118:1348-62
Lee, Mei Yan; Diamond, Scott L (2015) A human platelet calcium calculator trained by pairwise agonist scanning. PLoS Comput Biol 11:e1004118
Li, R; Panckeri, K A; Fogarty, P F et al. (2015) Recombinant factor VIIa enhances platelet deposition from flowing haemophilic blood but requires the contact pathway to promote fibrin deposition. Haemophilia 21:266-74
Muthard, Ryan W; Welsh, John D; Brass, Lawrence F et al. (2015) Fibrin, γ'-fibrinogen, and transclot pressure gradient control hemostatic clot growth during human blood flow over a collagen/tissue factor wound. Arterioscler Thromb Vasc Biol 35:645-54
Zhu, Shu; Herbig, Bradley A; Li, Ruizhi et al. (2015) In microfluidico: Recreating in vivo hemodynamics using miniaturized devices. Biorheology 52:303-18

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