Millions of platelet transfusions are conducted each year, yet the supply of this blood component is limited, thus patient access to treat disorders is problematic. There are also many diseases where platelet production or function are impaired, resulting in severe consequences and where there are limited clinical options available. To address these current limitations, new modes to generate functional platelets in vitro would provide a major benefit to many patients, as well as provide an approach to permit the systematic investigation of mechanisms involved in functional platelet formation. Our goal in this Project is to build upon our recent successful studies where a novel bioreactor system was engineered to house megakaryocytes (Mks) and to generate functional platelets in vitro. We will exploit this system to address our hypothesis;engineered microenvironments in vitro can be tailored to optimize the formation of functional platelets. To address the needs in the program we will:
(Aim 1) establish a bioreactor-based 3D tissue system to study the mechanisms of Mk development and platelet release, with functional outcomes in terms of functional platelets generated and recovered from the system, and (Aim 2) to use this 3D tissue system to investigate mechanisms of platelet production related to biochemical signalling and environmental components (i.e. matrices, oxygen tension) as well as pathological megakaryopoiesis by including Mks from patients with platelet related diseases. With insight from this system we will be positioned to interrogate the maturation of Mks from both normal vs. diseased sources in order to begin to establish differences in Mk outcomes (adhesion, migration, proplatelet formation, platelet production and function). The outcome for the proposed study would be twofold: (1) a new laboratory model for Mk development, proplatelet formation and platelet release in normal and abnormal (disease) states, and (2) mechanistic insight into these processes. In the long run, building upon this new in vitro tissue system would allow for the more systematic understanding of the processes involved in Mk development, as well as insight into modes to intervene in disease states associated with these cells.
Circulating platelets are highly specialized cells produced by Mks that participate in haemostatic and inflammatory functions. Millions of platelet transfusions are conducted each year but the supply of this blood component is limited. Despite their critical role in many physiological functions, little is known about the molecular mechanisms involved in platelet production from Mks, or about the pathogenesis of platelet disorders. With such widespread impact of these diseases, and the lack of good options for clinical treatments, new insight into the formation of platelets from Mks would have a major impact on patients and healthcare. To address this need, we plan to develop a novel human bone marrow system in vitro that can be used to understand the process of Mk formation and development both in normal and disease scenarios, as well as an innovative procedure to produce fully functional platelets for clinical needs.
|Abbonante, Vittorio; Di Buduo, Christian Andrea; Gruppi, Cristian et al. (2017) A new path to platelet production through matrix sensing. Haematologica 102:1150-1160|
|Balduini, Alessandra; Di Buduo, Christian A; Kaplan, David L (2016) Translational approaches to functional platelet production ex vivo. Thromb Haemost 115:250-6|
|Di Buduo, Christian A; Currao, Manuela; Pecci, Alessandro et al. (2016) Revealing eltrombopag's promotion of human megakaryopoiesis through AKT/ERK-dependent pathway activation. Haematologica 101:1479-1488|
|Abbonante, Vittorio; Di Buduo, Christian A; Gruppi, Cristian et al. (2016) Thrombopoietin/TGF-?1 Loop Regulates Megakaryocyte Extracellular Matrix Component Synthesis. Stem Cells 34:1123-33|
|Di Buduo, Christian A; Wray, Lindsay S; Tozzi, Lorenzo et al. (2015) Programmable 3D silk bone marrow niche for platelet generation ex vivo and modeling of megakaryopoiesis pathologies. Blood 125:2254-64|
|Malara, Alessandro; Abbonante, Vittorio; Di Buduo, Christian A et al. (2015) The secret life of a megakaryocyte: emerging roles in bone marrow homeostasis control. Cell Mol Life Sci 72:1517-36|
|Malara, Alessandro; Currao, Manuela; Gruppi, Cristian et al. (2014) Megakaryocytes contribute to the bone marrow-matrix environment by expressing fibronectin, type IV collagen, and laminin. Stem Cells 32:926-37|
|Pallotta, Isabella; Kluge, Jonathan A; Moreau, Jodie et al. (2014) Characteristics of platelet gels combined with silk. Biomaterials 35:3678-87|
|Rnjak-Kovacina, Jelena; Wray, Lindsay S; Golinski, Julianne M et al. (2014) Arrayed Hollow Channels in Silk-based Scaffolds Provide Functional Outcomes for Engineering Critically-sized Tissue Constructs. Adv Funct Mater 24:2188-2196|
|Di Buduo, Christian Andrea; Moccia, Francesco; Battiston, Monica et al. (2014) The importance of calcium in the regulation of megakaryocyte function. Haematologica 99:769-78|
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