Developing Novel Bispecific Antibody-Drug Conjugates to Treat Leukaemia

急性髓性白血病（AML）是第二常见的白血病，占成人中诊断的所有白血病的32%。AML是骨髓的侵袭性恶性肿瘤，其特征在于未分化的髓系细胞的不受控制的增殖和令人沮丧的存活率，特别是在老年患者中。AML给NHS带来了重大的临床负担，因为控制疾病所需的强化化疗方案往往会因副作用而导致住院治疗。因此，临床上需要一种新的治疗方法，以更特异的靶向治疗肿瘤，同时毒性最小。疾病复发也是AML中常见的问题，因为定义这种疾病的大部分恶性未分化髓系细胞来自一种罕见的称为白血病干细胞（LSC）的祖细胞群体，并由其维持。LSC具有自我更新、长期克隆繁殖和分化的能力，但正是它们的静止能力为它们提供了对一线疗法的抗性。因此，LSC的特异性去除在AML的治疗中是明显的优势，并且可能使用新型双特异性抗体药物缀合物（ADC）来实现，所述新型双特异性抗体药物缀合物能够通过结合限定的表面抗原并通过有毒货物的内化来识别并杀死这些细胞。导致恶性髓系细胞形成AML。与我们的工业合作伙伴BiVictriX一起，我们计划专注于鉴定、开发和测试针对AML中LSC的新型抗体-药物缀合物（ADC）。实验方法我们的实验方法将利用免疫学、细胞生物学、分子生物学、生物技术领域的最先进技术以及先进的数据分析方法。首先，我们将使用质量细胞术（CyTOF），这是一种利用“重金属标记探针”的新技术，允许测量每个细胞的>40个表型属性，以提供基础数据，这将有助于加速识别新的抗原组合，为ADC靶点的Biviclets开发管道提供信息。来自AML和其他患者的细胞群的比较将允许鉴定表面抗原表型，例如但不限于表达的⑶ 34、⑶ 38、⑶ 133和⑶ 90，其将LSC与非恶性对应细胞区分开。我们还打算通过以下组合来功能性表征这些新型ADC的功效：（i）先进的显微镜以研究ADC在细胞系模型中的摄取和加工;（ii）一系列细胞毒性测定，其测试ADC在表达适当抗原的细胞系模型中的特异性和选择性;（iii）最终确定体外患者材料中的有效性。

Acute myeloid leukaemia (AML) is the second most common leukaemia, accounting for 32% of all leukaemia in diagnosed in adults. AML is an aggressive malignancy of the bone marrow, as is characterized by uncontrolled proliferation of undifferentiated myeloid lineage cells and dismal survival rates, particularly among older patients. AML presents a significant clinical burden to the NHS because the intensive chemotherapy regimens necessary to bring the disease under control often result in hospitalisation due to side effects. There is therefore distinct clinical need of new therapies that will more specifically target the tumour with minimal toxicity.Disease relapse is also a problem that is common in AML because the bulk of malignant undifferentiated myeloid lineage cells that define this disease arise from, and are sustained by, a rare population of progenitor cells called leukemic stem cells (LSC). LSCs have the capacity for self-renewal, long-term clonal propagation and for differentiation, but it is their capacity for quiescence that provides them with resistance to front line therapies. Specific removal of LSCs would therefore be a distinct advantage in the therapy of AML, and could potentially be achieved using novel bi-specific antibody drug conjugates (ADCs) that are able to recognise and kill these cells by binding defined surface antigens and through internalisation of a toxic cargo.Objectives.Our principle goal is to remove the leukemic stem cells (LSC) that give rise to the malignant myeloid lineage cells that define AML. With our industrial partner, BiVictriX, we plan to focus on identification, development and testing of novel antibody-drug conjugates (ADCs) that target LSCs in AML.Experimental approach.Our experimental approach will utilize state-of-the-art techniques in covering areas of immunology, cell biology, molecular biology, biotechnology alongside advanced data analysis approaches. Primarily we will use mass cytometry (CyTOF), a novel technology that utilises 'heavy metal tagged probes' allowing measurement of >40 phenotypic attributes per cell, to provide foundation data that will help to accelerate identification of novel combinations of antigens to inform the Bivictrix development pipeline of ADC targets. Comparisons of cell populations derived from AML and other patients will allow identification of surface antigen phenotypes, such as but not restricted to expressed CD34, CD38, CD133 and CD90, that differentiate LSCs from non-malignant counterpart cells. We also intend to functionally characterize the efficacy of these novel ADCs through a combination of (i) advanced microspcopy to study uptake and processing of the ADC in cell line models; (ii) a range of cytototoxicity assays testing the specificity and selectivity of the ADCs in cell line models expressing appropriate antigens; (iii) finally to determine the effectiveness in patient material in vitro.