Development and validation of 3D in vitro dormant myeloma cell models to reduce and replace animal studies

开发和验证 3D 体外休眠骨髓瘤细胞模型，以减少和替代动物研究

• 批准号: 2594640
• 金额: --
• 依托单位: University of Sheffield
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2594640
• 关键词: Development; validation; 3D; vitro; dormant

Scientific importance: Multiple myeloma (MM) is a B cell cancer caused by the growth of malignant plasma cells in the bone marrow (BM). It remains incurable, despite the development of therapies that target myeloma cell (MC)-intrinsic pathways. Disease relapse is thought to originate from dormant MCs, localised in specialised endosteal niches, which resist chemotherapy and repopulate the tumour [1, 2]. We have previously tracked individual MC's in vivo by multi-photon imaging as they colonise the endosteal niche, enter a dormant state and subsequently become activated to form colonies [1]. MC dormancy has been demonstrated to be a reversible state that is switched 'on' by engagement with osteoblasts (in the endosteal niche) and 'off' by osteoclastic bone resorption. We have determined dormant MCs are resistant to the chemotherapy melphalan that targets dividing cells. The demonstration that the endosteal niche is pivotal in controlling MC dormancy highlights the potential for targeting cell-extrinsic mechanisms to overcome cell-intrinsic drug resistance and prevent disease relapse. To recapitulate the BM microenvironment in an in vitro setting is highly challenging because of the cellular complexity, vasculature and 3-dimensional (3D) bone structure. However, since we know osteoblasts play a key role in MC dormancy and they reside on bone surfaces in close proximity to the extracellular matrix (ECM), this will provide a platform to mimic this in vivo aspect in vitro. Therefore, the aim of this 3Rs-focussed PhD studentship is to develop several in vitro 3D models of MC dormancy and then validate them by comparing them to new data and our existing data from established clinically relevant murine models of MC dormancy and drug resistance.Research plan and objectives: We aim to develop and validate several 3D in vitro models of MC dormancy which recapitulates the interaction of MC-osteoblastic cells in the BM microenvironment in vivo i.e. induces MC dormancy. We will do this by assessing and characterising dormant MCs in these newly established 3D in vitro models so they can be used reliability as effective alternatives to murine models of myeloma to assess new and novel anti-myeloma therapies and their mechanisms of action. We hypothesise that in vitro 3D models of MC dormancy will mimic the dormant MC's observed in the endosteal niche in the BM of currently established in vivo pre-clinical models of myeloma.

多发性骨髓瘤（MM）是一种由骨髓（BM）中恶性浆细胞生长引起的B细胞癌。尽管针对骨髓瘤细胞（MC）内在途径的治疗方法不断发展，但它仍然无法治愈。疾病复发被认为起源于休眠的MC，定位于专门的骨内膜龛，其抵抗化疗并重新填充肿瘤[1，2]。我们之前已经通过多光子成像在体内追踪了单个MC，因为它们定植在骨内膜龛中，进入休眠状态，随后被激活形成菌落[1]。MC休眠已被证明是一种可逆的状态，通过与成骨细胞（在骨内膜龛中）的接合而“打开”，并通过骨细胞骨吸收而“关闭”。我们已经确定休眠的MC对针对分裂细胞的化疗美法仑具有抗性。骨内膜生态位在控制MC休眠中起关键作用的证明突出了靶向细胞外在机制以克服细胞内在耐药性和预防疾病复发的潜力。由于细胞复杂性、脉管系统和三维（3D）骨结构，在体外环境中概括BM微环境极具挑战性。然而，由于我们知道成骨细胞在MC休眠中起着关键作用，并且它们位于骨表面上靠近细胞外基质（ECM），因此这将提供一个在体外模拟体内方面的平台。因此，这个以3RS为重点的博士生项目的目的是开发几种MC休眠的体外3D模型，然后通过将它们与新数据和我们现有的MC休眠和耐药性临床相关小鼠模型的数据进行比较来验证它们。研究计划和目标：我们的目标是开发和验证几种MC休眠的三维体外模型，该模型概括了MC-成骨细胞在体内的BM微环境中，即诱导MC休眠。我们将通过评估和表征这些新建立的3D体外模型中的休眠MC来实现这一点，因此它们可以可靠地用作骨髓瘤小鼠模型的有效替代品，以评估新的和新颖的抗骨髓瘤疗法及其作用机制。我们假设MC休眠的体外3D模型将模拟目前建立的骨髓瘤体内临床前模型的BM中骨内膜龛中观察到的休眠MC。