Miniaturized 3D bone marrow tissue model to assess response to Thrombopoietin-receptor agonists in patients.
Miniaturized 3D bone marrow tissue model to assess response to Thrombopoietin-receptor agonists in patients.
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DOI：
10.7554/elife.58775
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发表时间：
2021-06-01
期刊：
影响因子：
7.7
通讯作者：
Balduini A
中科院分区：
文献类型：
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作者：
Di Buduo CA;Laurent PA;Zaninetti C;Lordier L;Soprano PM;Ntai A;Barozzi S;La Spada A;Biunno I;Raslova H;Bussel JB;Kaplan DL;Balduini CL;Pecci A;Balduini A
血小板减少性疾病已使用血小板生成素受体激动剂艾曲泊帕治疗。具有相同血小板减少症表观形式的患者可能对治疗有不同的反应。我们描述了一种小型化骨髓组织模型，该模型提供了一种筛选生物反应器，用于个体患者对艾曲泊帕的个性化治疗前反应预测。使用丝素蛋白，开发了一个3D骨髓龛，再现血小板生物合成。从ANKRD 26和MYH 9基因突变患者的少量外周血中分离造血祖细胞，这些患者既往接受过艾曲泊帕治疗。离体反应与体内血小板反应密切相关。1例MYH 9突变患者的诱导多能干细胞（iPSC）分化为对艾曲泊帕有反应的功能性巨核细胞。将患者来源的细胞和iPSC与3D骨髓模型技术相结合，可以拥有一个可重复的系统，用于研究药物机制，并用于遗传性血小板减少症的有效治疗的个体化治疗前选择。血小板是血液凝结所必需的微小细胞碎片。它们由巨核细胞产生并释放到血液中，巨核细胞是生活在骨髓中的巨细胞。在某些遗传性疾病中，如遗传性血小板减少症，骨髓无法产生足够的血小板：这使得患者在受伤或手术后极易出现瘀伤，出血和凝血不良。某些遗传性血小板减少症患者对旨在促进血小板生成的治疗反应良好，但其他患者则不然。这些差异存在的原因可以通过设计新的测试系统来研究，这些测试系统可以在实验室中重现骨髓的形式和功能。然而，在体外构建复杂且知之甚少的骨髓环境是具有挑战性的。在这里，狄布多等人开发了一种人造三维微型器官生物反应器系统，重现了骨髓的关键特征。在这个系统中，巨核细胞从病人的血液样本中生长出来，并连接到由丝制成的组织支架上。这些细胞能够像在正常环境中一样生长，它们可以将血小板释放到人造血液中。在用刺激血小板生成的药物处理巨核细胞后，迪布多等人发现，从生物反应器中回收的血小板数量可以准确预测哪些患者在临床上会对这些药物产生反应。这种新的测试系统使研究人员能够预测患者对治疗的反应，并为每个人量身定制治疗方案。这项技术也可以用于测试遗传性血小板减少症和其他血液相关疾病的新药;如果扩大规模，它也可以在某一天产生大量的实验室培养的血细胞用于输血。
Thrombocytopenic disorders have been treated with the Thrombopoietin-receptor agonist Eltrombopag. Patients with the same apparent form of thrombocytopenia may respond differently to the treatment. We describe a miniaturized bone marrow tissue model that provides a screening bioreactor for personalized, pre-treatment response prediction to Eltrombopag for individual patients. Using silk fibroin, a 3D bone marrow niche was developed that reproduces platelet biogenesis. Hematopoietic progenitors were isolated from a small amount of peripheral blood of patients with mutations in ANKRD26 and MYH9 genes, who had previously received Eltrombopag. The ex vivo response was strongly correlated with the in vivo platelet response. Induced Pluripotent Stem Cells (iPSCs) from one patient with mutated MYH9 differentiated into functional megakaryocytes that responded to Eltrombopag. Combining patient-derived cells and iPSCs with the 3D bone marrow model technology allows having a reproducible system for studying drug mechanisms and for individualized, pre-treatment selection of effective therapy in Inherited Thrombocytopenias. Platelets are tiny cell fragments essential for blood to clot. They are created and released into the bloodstream by megakaryocytes, giant cells that live in the bone marrow. In certain genetic diseases, such as Inherited Thrombocytopenia, the bone marrow fails to produce enough platelets: this leaves patients extremely susceptible to bruising, bleeding, and poor clotting after an injury or surgery. Certain patients with Inherited Thrombocytopenia respond well to treatments designed to boost platelet production, but others do not. Why these differences exist could be investigated by designing new test systems that recreate the form and function of bone marrow in the laboratory. However, it is challenging to build the complex and poorly understood bone marrow environment outside of the body. Here, Di Buduo et al. have developed an artificial three-dimensional miniature organ bioreactor system that recreates the key features of bone marrow. In this system, megakaryocytes were grown from patient blood samples, and hooked up to a tissue scaffold made of silk. The cells were able to grow as if they were in their normal environment, and they could shed platelets into an artificial bloodstream. After treating megakaryocytes with drugs to stimulate platelet production, Di Buduo et al. found that the number of platelets recovered from the bioreactor could accurately predict which patients would respond to these drugs in the clinic. This new test system enables researchers to predict how a patient will respond to treatment, and to tailor therapy options to each individual. This technology could also be used to test new drugs for Inherited Thrombocytopenias and other blood-related diseases; if scaled-up, it could also, one day, generate large quantities of lab-grown blood cells for transfusion.