NUOVE STRATEGIE PER LO STUDIO DELLA PRODUZIONE PIASTRINICA
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1 NUOVE STRATEGIE PER LO STUDIO DELLA PRODUZIONE PIASTRINICA Alessandra Balduini, MD University of Pavia (Pavia, Italy) Tufts University (Boston, MA, USA) Milano,
2 PLATELET FORMATION Balduini et al., JTH 2008 PLATELET PRODUCTION Alessandra Balduini Milano,
3 MK DISTRIBUTION IN BONE MARROW Malara et al., Stem Cells in press PLATELET PRODUCTION Alessandra Balduini Milano,
4 ECMs IN BONE MARROW NICHES Malara et al., Stem Cells in press PLATELET PRODUCTION Alessandra Balduini Milano,
5 PROPLATELET REGULATION MODEL Chen et al., Blood 2007 PLATELET PRODUCTION Alessandra Balduini Milano,
6 PROPLATELET RELEASE IN MYH9 PATIENT DERIVED MKS Control MYH9-patients Control BSA 1/A BSA 2/B BSA 3/C BSA 3/C Col-I Pecci et al., TH 2009 PLATELET PRODUCTION Alessandra Balduini Milano,
7 MAMMALIAN COLLAGEN RECEPTORS PLATELET PRODUCTION Alessandra Balduini Milano,
8 DISCOIDIN DOMAIN RECEPTOR 1 Abbonante et al., JBC 2013 PLATELET PRODUCTION Alessandra Balduini Milano,
9 DISCOIDIN DOMAIN RECEPTOR 1 AND SYK Abbonante et al., JBC 2013 PLATELET PRODUCTION Alessandra Balduini Milano,
10 DISCOIDIN DOMAIN RECEPTOR 1 AND SYK Abbonante et al., JBC 2013 PLATELET PRODUCTION Alessandra Balduini Milano,
11 DISCOIDIN DOMAIN RECEPTOR 1 AND SHP1 Abbonante et al., JBC 2013 PLATELET PRODUCTION Alessandra Balduini Milano,
12 - Phosphatase SHP1 resulted less phosphorilated upon blocking DDR1 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge, SUMMARY -ECMs are differently located in bone marrow niches, further supporting their involvement in the regulation of hematopoiesis. -A gradients of maturing MKs characterizes the vascular niche in the bone marrow. -We identified the expression of a functional new collagen receptor in human Mk (DDR1) and provided evidence indicating DDR1 implication in Mks migration -Migration inhibitor Syk kinase resulted more phosphorilated upon blocking DDR1 activation
13 FIBRONECTIN MODULATES MK ADHESION ON ECM Malara et al., Blood 2011 PLATELET PRODUCTION Alessandra Balduini Milano,
14 MODEL OF MK INTERACTION WITHIN BONE MARROW ENVIRONMENT Malara et al., Blood 2011 PLATELET PRODUCTION Alessandra Balduini Milano,
15 FIBRONECTIN MODULATES MK ADHESION ON ECM Malara et al., Stem Cells in press PLATELET PRODUCTION Alessandra Balduini Milano,
16 TYPE I COLLAGEN STRUCTURE Malara et al., Blood 2012 PLATELET PRODUCTION Alessandra Balduini Milano,
17 NANO-MECHANICAL PROPERTIES Malara et al., Blood 2012 PLATELET PRODUCTION Alessandra Balduini Milano,
18 NANO-MECHANICAL PROPERTIES Type I collagen N-acetylated Type I collagen Malara et al., Blood 2012 PLATELET PRODUCTION Alessandra Balduini Milano,
19 N-acetylated type I collagen Type I collagen FIBRONECTIN ASSEMBLY Cellular Fibronectin Plasma Fibronectin Malara et al., Blood 2012 PLATELET PRODUCTION Alessandra Balduini Milano,
20 IMPACT OF COLLAGEN NANO-MECHANICS ON pmlc2 Malara et al., Blood 2012 PLATELET PRODUCTION Alessandra Balduini Milano,
21 TYPE IV COLLAGEN TYPE I COLLAGEN ROLE OF DIFFERENT ECMs ON PLATELET RELEASE Balduini et al., JTH 2008 PLATELET PRODUCTION Alessandra Balduini Milano,
22 ECMs IN BONE MARROW NICHES Malara et al., Stem Cells in press PLATELET PRODUCTION Alessandra Balduini Milano,
23 ECMS DIFFERENTLY REGULATE MK DEVELOPMENT Malara et al., Stem Cells in press PLATELET PRODUCTION Alessandra Balduini Milano,
24 ECMs IN BONE MARROW NICHES Malara et al., Stem Cells in press PLATELET PRODUCTION Alessandra Balduini Milano,
25 ECM EXPRESSION BY MKs Malara et al., Stem Cells in press PLATELET PRODUCTION Alessandra Balduini Milano,
26 MYELOSUPPRESSION AND PLATELET DEPLATION Malara et al., Stem Cells in press PLATELET PRODUCTION Alessandra Balduini Milano,
27 MYELOSUPPRESSION AND PLATELET DEPLATION Malara et al., Stem Cells in press PLATELET PRODUCTION Alessandra Balduini Milano,
28 MK SORTING Malara et al., Stem Cells in press PLATELET PRODUCTION Alessandra Balduini Milano,
29 ECM EXPRESSION BY MKs Malara et al., Stem Cells in press PLATELET PRODUCTION Alessandra Balduini Milano,
30 SUMMARY -Fibronectin and factor XIII regulate Mk interaction with type I collagen -Collagen biomechanics impact Mk function -Fibronectin, type IV collagen and Laminin differently regulate Mk development -Mks express laminin, type IV collagen and fibronectin -Upon 5-FU treatment Mks contribute to the restoration of bone marrow environment by over-expressing laminin, type IV collagen and fibronectin. -Upon platelet depletion the over-expression of Mk derived ECMs is restricted to type IV collagen PLATELET PRODUCTION Alessandra Balduini Milano,
31 P2Y EXPRESSION Balduini et al. Haematologica 2012 PLATELET PRODUCTION Alessandra Balduini Milano,
32 EFFECT OF SPECIFIC P2Y13 INHIBITION Balduini et al. Haematologica 2012 PLATELET PRODUCTION Alessandra Balduini Milano,
33 TGFβ1 RECEPTORS ARE EXPRESSED BY HUMAN MKS Badalucco et al. Haematologica 2013 PLATELET PRODUCTION Alessandra Balduini Milano,
34 Akt AND REGULATION OF PLATELET RELEASE ADP TGFβ1 PLATELET PRODUCTION Alessandra Balduini Milano,
35 HISTORICAL PATIENTS WITH BLEEDING DIATHESIS ( ) Balduini et al. Haematologica 2012 PLATELET PRODUCTION Alessandra Balduini Milano,
36 PMF-DERIVED MK CULTURES Balduini et al. PLoS ONE 2011 PLATELET PRODUCTION Alessandra Balduini Milano,
37 SUMMARY -Autocrine components seem to play a crucial role in the regulation of platelet production. - ADP is released by human megakaryocytes and regulates proplatelet formation through P2Y13 and Akt activation. - TGFβ1 is released by human megakaryocytes and regulates proplatelet formation through SMAD2/3 and Akt activation. PLATELET PRODUCTION Alessandra Balduini Milano,
38 Silk Silk fibers
39 SILK AND BIOMEDICAL RESEARCH Tubes: Silk Extract Microparticles: Electrospun fibers: Porous scaffolds: Omenetto et al., Science 2010 PLATELET PRODUCTION Alessandra Balduini Milano,
40 SILK AND IS AN INERT MATERIAL FOR PLATELETS WBC 10 3 /µl NEU 10 3 /µl LYM 10 3 /µl MONO 10 3 /µl EOS 10 3 /µl BASO 10 3 /µl RBC 10 6 /µl HGB g/dl HCT % MCV fl MCH pg MCHC g/dl RDW % PLT 10 3 /µl Before perfusion After perfusion PLATELET PRODUCTION Alessandra Balduini Milano,
41 NEW OPPORTUNITIES FOR AN ANCIENT MATERIAL Omenetto et al., Science 2010; Hwang et al., Science 201 PLATELET PRODUCTION Alessandra Balduini Milano,
42 TUBE MODEL AND MKs Wray et al., in prparation PLATELET PRODUCTION Alessandra Balduini Milano,
43 TUBE MODEL AND MKs Wray et al., in prparation PLATELET PRODUCTION Alessandra Balduini Milano,
44 SUMMARY We started including general protein components in the bioreactor model, then, after the study of the bone marrow in vivo, we created a second model with more physiologic characteristics. PLATELET PRODUCTION Alessandra Balduini Milano,
45 ACKNOLEDGEMENTS University of Pavia-Tufts University Alessandro Malara Manuela Currao Cristian Gruppi Vittorio Abbonante Christian Di Buduo Lindsay Wray Lorenzo Tozzi Ying Chen Bentley Hunt Collaborators Luigi De Marco Maria Enrica Tira David L. Kaplan Cristian Staii Giovanni Barosi Vittorio Rosti Carlo Balduini Alessandro Pecci
46 HUMAN MK BEHAVIOR ON MATRIX COMPONENTS Di Buduo et al., submitted BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
47 UNDERSTANDING DISEASES MYH9-RD ET Pecci et al.th 2009; Balduini et al., JTH 2009; Noris et al., Platelets 2010; Vettore et al., Haematologica 2011; Balduini et al., Blood 2011; Balduini et al., PLoS ONE 2011; Currao et al. PLoS ONE 2013; Necchi et al., TH 2013; Badalucco et al., Haematologica 2013 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
48 THROMBOCYTOPENIA IN DELTA-SPD Study Thrombocytopenias/total Weiss HJ, BLOOD /18 (6%) Nieuwenhuis HK, BLOOD /51 (18%) Pujol-Moix N, HAEMATOLOGICA /24 (42%) Balduini et al. Haematologica 2012 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
49 ADP AND PLATELET FORMATION Balduini et al. Haematologica 2012 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
50 ADP AND PLATELET FORMATION Balduini et al. Haematologica 2012 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
51 P2Y13 SIGNALING IN MKs Balduini et al. Haematologica 2012 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
52 TGFβ1 IS RELESED DURING MK MATURATION * * Badalucco et al. Haematologica 2013 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
53 TGFβ1 AND PROPLATELET FORMING MKS *p<0.05 Badalucco et al. Haematologica 2013 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
54 PMF-DERIVED MK CULTURES Balduini et al. PLoS ONE 2011 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
55 TGF 1 IN PMF-DERIVED MKS CULTURES Badalucco et al. Haematologica 2013 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
56 SILK VASCULAR TUBES BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
57 3D BONE MARROW MODEL USING SILK BASED VASCULAR TUBES Pallotta et al.tissue Engineering 2011 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
58 RELEASED PLATELETS IN 3D MODEL Pallotta et al.te 2011 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
59 RELEASED PLATELETS IN 3D MODEL Pallotta et al.te 2011 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
60 SUMMARY BONE MARROW MODELS -Silk-based model. Alessandra Balduini. Tufts and University of Pavia. GRC Tissue Eng Part C Methods; 17(12): Collagen-based bioreactor. Cedric Gevahert. Cambridge. GRC PDMS-based bioreactor. Joseph Italiano. Harvard. GRC Collagen-gel bioreactor. Jose Lopez. University of Washington School of Medicine. ISTH PDMS-based bioreactor. William Miller. Northwestern University. GRC ips and PDMS based-bioreactor. Koji Eto. Kyoto. Exp Hematol Aug;41(8): BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
61 SILK AND BIOMEDICAL RESEARCH Wray et al., in prparation BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
62 SILK AND BIOMEDICAL RESEARCH Wray et al., in prparation BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
63 PROPLATELET REGULATION MODEL Malara et al., submitted BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
64 RELEASED PLATELETS IN 3D MODEL
65 SILK AND BIOMEDICAL RESEARCH Wray et al., in prparation BM ENVIRONMENT AND MKs Alessandra Balduini Sperlonga,
66
67 Tube Model sponge tube Sponge Tube wall Tube wall Inner tube wall Tube lumen 2
68 TUBE MODEL BM ENVIRONMENT AND MKs Alessandra Balduini Sperlonga,
69 Silk and the Vascular Niche Tube Model: Membrane Model:
70 Silk and Biomedical Research Tubes: Silk Extract Microparticles: Electrospun fibers: Porous scaffolds: Silk Films:
71 ECMs IN BONE MARROW NICHES Malara et al., submitted MEGACARIOCITI E PROPIASTRINE Alessandra Balduini Aviano,
72 PRIMARY MYELOFIBROSIS (PMF) clonality of hematopoiesis: proliferation of myeloid and megakaryocytic lines marrow fibrosis: fibrotic cytokines and ECM accumulation extramedullary hematopoiesis: enhanced cellular trafficking of CD34+ in PB Histological features: - Hyperplasia of megakaryocytic line - Small, aggregated, naked and cloud-like nuclei Reilly, J Clin Pathol 1992 ECM AND STEM CELL REGULATION Alessandra Balduini Pavia,
73 OF MEN, NOT MICE (NATURE MEDICINE (2013), 19, 379) Pallotta et al.te 2011 MEGACARIOCITI E PROPIASTRINE Alessandra Balduini Aviano,
74 NEW OPPORTUNITIES FOR AN ANCIENT MATERIAL Omenetto et al., Science 2010; Hwang et al., Science 201 MEGACARIOCITI E PROPIASTRINE Alessandra Balduini Aviano,
75 PROPLATELET REGULATION MODEL Chen et al., Blood 2007 MEGACARIOCITI E PROPIASTRINE Alessandra Balduini Aviano,
76 MEGAKARYOCYTES EXPRESS FIBRONECTIN Malara et al., Blood 2011 MEGACARIOCITI E PROPIASTRINE Alessandra Balduini Aviano,
77 FIBRONECTIN MODULATES MK ADHESION ON ECM Malara et al., Blood 2011 MEGACARIOCITI E PROPIASTRINE Alessandra Balduini Aviano,
78 FIBRONECTIN AND FXIII-A MODULATE MK ADHESION ON ECM Malara et al., Blood 2011 MEGACARIOCITI E PROPIASTRINE Alessandra Balduini Aviano,
79 MODEL OF MK INTERACTION WITHIN BONE MARROW ENVIRONMENT Malara et al., Blood 2011 MEGACARIOCITI E PROPIASTRINE Alessandra Balduini Aviano,
80 FIBRONECTIN MODULATES MK ADHESION ON ECM Malara et al., submitted MEGACARIOCITI E PROPIASTRINE Alessandra Balduini Aviano,
81 ECMs IN BONE MARROW NICHES Malara et al., submitted MEGACARIOCITI E PROPIASTRINE Alessandra Balduini Aviano,
82 ECMs IN BONE MARROW NICHES Malara et al., submitted MEGACARIOCITI E PROPIASTRINE Alessandra Balduini Aviano,
83 ECMs IN BONE MARROW NICHES Malara et al., submitted MEGACARIOCITI E PROPIASTRINE Alessandra Balduini Aviano,
84 MAMMALIAN COLLAGEN RECEPTORS MEGACARIOCITI E PROPIASTRINE Alessandra Balduini Aviano,
85 DISCOIDIN DOMAIN RECEPTOR 1 Abbonante et al., JBC in press MEGACARIOCITI E PROPIASTRINE Alessandra Balduini Aviano,
86 ABNORMAL MK BEHAVIOR ON MATRIX COMPONENTS CONTROL Controls PATIENTS (REGULAR SPREADING) 20 μm 20 μm PATIENTS (ABNORMAL SPREADING) PATIENTS (ABNORMAL SPREADING) 20 μm 20 μm Bury et al., PLoS ONE 2012 MEGACARIOCITI E PROPIASTRINE Alessandra Balduini Aviano,
87 UNDERSTANDING DISEASES MYH9-RD ET Pecci et al.th 2009; Balduini et al., JTH 2009; Noris et al., Platelets 2010; Vettore et al., Haematologica 2011; Balduini et al., Blood 2011; Balduini et al., PLoS ONE 2011; Currao et al. PLoS ONE 2013; Necchi et al., TH 2013; Badalucco et al., Haematologica 2013 MEGACARIOCITI E PROPIASTRINE Alessandra Balduini Aviano,
88 ANOIKIS: INTRINSIC AND EXTRINSIC PATHWAYS ECM AND STEM CELL REGULATION Alessandra Balduini Pavia,
89 ANOIKIS RESISTANCE ECM AND STEM CELL REGULATION Alessandra Balduini Pavia,
90 ANOIKIS RESISTANCE ECM AND STEM CELL REGULATION Alessandra Balduini Pavia,
91 ECMs IN BONE MARROW NICHES Malara et al., submitted ECM AND STEM CELL REGULATION Alessandra Balduini Pavia,
92 PROPLATELET RELEASE IN MYH9 PATIENT DERIVED MKS Control MYH9-patients Control BSA 1/A BSA 2/B BSA 3/C BSA 3/C Col-I Pecci et al., TH 2009 ECM AND STEM CELL REGULATION Alessandra Balduini Pavia,
93 ABNORMAL MK BEHAVIOR ON MATRIX COMPONENTS Bury et al., PLoS ONE 2012 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
94 % of PPF ABNORMAL MK BEHAVIOR ON MATRIX COMPONENTS CONTROLS PATIENTS Controls Patients Suspension FBG Coll I VWF Bury et al., PLoS ONE 2012 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
95 BERNARD SOULIER BOLZANO Balduini et al., JTH 2009 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
96 BERNARD SOULIER BOLZANO Balduini et al., JTH 2009 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
97 DISCOIDIN DOMAIN RECEPTOR 1 AND SYK Abbonante et al., in revision BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
98 DISCOIDIN DOMAIN RECEPTOR 1 AND SYK Abbonante et al., in revision BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
99 DISCOIDIN DOMAIN RECEPTOR 1 AND SHP1 Abbonante et al., in revision BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
100 ROMIPLOSTIM AND MK FUNCTION Currao et al., PLoS ONE 2013 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
101 RECOMBINANT TPO SIGNALING Currao et al., PLoS ONE 2013 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
102 ADP ELICITS CA2+ MOBILIZATION IN HUMAN MEGAKARYOCYTES Di Buduo et al. submitted BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
103 ADP-INDUCED SUSTAINED ELEVATION IN [CA2+]I REQUIRES CA2+ ENTRY Di Buduo et al. submitted BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
104 STORE-OPERATED CA2+ ENTRY (SOCE) IN MKS Di Buduo et al. submitted BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
105 ADP-INDUCED Ca2+ MOBILIZATION DOWNSTREAM SIGNALING BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
106 CA2+ SIGNALING IMPACT ON MK ADHESION AND PROPLATELE FORMATION Di Buduo et al. submitted BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
107 CA2+ SIGNALING IMPACT ON MK MIGRATION Di Buduo et al. submitted BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
108 SILK BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
109 NEW OPPORTUNITIES FOR AN ANCIENT MATERIAL Omenetto et al., Science 2010; Hwang et al., Science 201 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
110 NEW OPPORTUNITIES FOR AN ANCIENT MATERIAL Omenetto et al., Science 2010; Hwang et al., Science 201 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
111 SILK VASCULAR TUBES BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
112 STORE-OPERATED CA2+ ENTRY (SOCE) IN MKS Di Buduo et al. submitted BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
113 CA2+ SIGNALING IMPACT ON MK MIGRATION Di Buduo et al. submitted BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
114 EXPRESSION PROFILING OF ESC DERIVED MKS Merico et al., JTH 2012 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
115 CONCLUSIONS Il vero viaggio di scoperta non consiste nel cercare nuove terre, ma nell'avere nuovi occhi. Marcel Proust BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
116 % of PPF Controls Patients CONTROLS PATIENTS Suspension FBG Coll I VWF
117
118 P2Y13 SIGNALING IN MKs Balduini et al. Haematologica 2012 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
119 Adapted from Rich JN, Fron Biosci TGFβ1 receptors are expressed by human Mks and the related signaling pathways are activated
120 Conclusions Autocrine components seem to play a crucial role in the regulation of platelet production. TGFβ1 is released by human megakaryocytes and regulates proplatelet formation through SMAD2/3 and Akt activation. Increased levels of released bioactive TGFβ1 by PMF-derived megakaryocytes did not enhance TGFβ1 receptor downstream signaling in the latest stages of megakaryocyte differentiation. The aberrant megakaryopoiesis that occurs in the fibrotic bone marrow of PMF patients could be due to an excess of TGFβ1 that probably affects the first stages of cell differentiation rather than proplatelet formation.
121 UNDERSTANDING DISEASES Pecci et al.th 2009; Balduini et al., JTH 2009; Noris et al., Platelets 2010; Vettore et al., Haematologica 2011; Balduini et al., Blood 2011; Balduini et al., PLoS ONE 2011; Currao et al. PLoS ONE submitted; Necchi et al., TH submitted; Badalucco et al., Haematologica submitted BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini - Chieti,
122 SUMMARY OF TECHNIQUES Human and mouse models (control and disease models) Cell Biology (human, cord blood, mouse, ES, ips) Molecular Biology and Biochemistry Flow cytometry and IF microscopy Atomic force microscopy Calcium analysis and membrane potential Silk processing, bioreactors, oxygen tension FUNCTIONAL PLATELET PRODUCTION Alessandra Balduini, MD - Boston,
123 PLATELET CONCENTRATES About 13 million platelet concentrates are collected annually in Europe (10 million in USA) at a cost of about 0.75 billion-dollars. They are needed by people who lack platelets or whose platelets function improperly, such as certain cancer chemotherapy, bone marrow transplant patients, trauma patients given massive blood transfusion and people with inherited thrombocytopenias. The short shelf life means that platelets cannot not easily be shipped from an area of surplus to one of scarcity, and hospitals occasionally experience shortages that require surgeries to be postponed. FUNCTIONAL PLATELET PRODUCTION Alessandra Balduini, MD - Boston,
124 ROLE OF DIFFERENT ECMs ON MK DEVELOPMENT Malara et al., submitted BONE MARROW AND PLATELET PRODUCTION BONE MARROW ANAlessandra Balduini NYC,
125 ROLE OF DIFFERENT ECMs ON PLATELET RELEASE Malara et al., submitted BONE MARROW AND PLATELET PRODUCTION BONE MARROW ANAlessandra Balduini NYC,
126 ROLE OF ADP IN REGULATING MK-MATRIX INTERACTION Di Buduo et al. submitted BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
127 STORE-OPERATED CA2+ ENTRY (SOCE) IN MKS Di Buduo et al. submitted BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
128 SOCE IS ENGAGED BY ADP STIMULATION IN HUMAN MKS Di Buduo et al. submitted
129 STUDIES OF ADP-INDUCED CALCIUM SIGNALING IN MKS Di Buduo et al. submitted BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
130 PMF-DERIVED MK CULTURES MEGAKARYOCYTES OUTPUT * * p<0.05 CTRL: median 2.2% (range ) PRE-PMF: median 7.2% (range ) ET: median 8.3% (range ) Di Buduo et al., submitted ECM AND STEM CELL REGULATION Alessandra Balduini Pavia,
131 PMF-DERIVED MK CULTURES PROPLATELET FORMATION * * * p<0.05 CTRL: median 7.4% (range 7 11) PRE-PMF: median 3.7% (range 1 8.9) ET: median 10.1% (range ) Di Buduo et al., submitted ECM AND STEM CELL REGULATION Alessandra Balduini Pavia,
132 STUDIES OF ADP-INDUCED CALCIUM SIGNALING IN MKS Di Buduo et al. submitted BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini NYC,
133 ABNORMAL MK ON MATRIX COMPONENTS CONTROL Controls PATIENTS (REGULAR SPREADING) 20 μm 20 μm PATIENTS (ABNORMAL SPREADING) PATIENTS (ABNORMAL SPREADING) 20 μm 20 μm Bury et al., PLoS ONE 2012 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
134 ABNORMAL MK BEHAVIOR ON MATRIX COMPONENTS Bury et al., PLoS ONE 2012 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
135 % of PPF ABNORMAL MK BEHAVIOR ON MATRIX COMPONENTS CONTROLS PATIENTS Controls Patients Suspension FBG Coll I VWF Bury et al., PLoS ONE 2012 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
136 BERNARD SOULIER BOLZANO Balduini et al., JTH 2009 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
137 BERNARD SOULIER BOLZANO Balduini et al., JTH 2009 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
138 MEGAKARYOCYTES EXPRESS FIBRONECTIN Malara et al., Blood 2011 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
139 FIBRONECTIN AND FXIII-A MODULATE MK ADHESION ON ECM Malara et al., Blood 2011 BONE MARROW AND PLATELET PRODUCTION Alessandra Balduini Cambridge,
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