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，占成人诊断的所有白血病的32%。AML是一种侵袭性骨髓恶性肿瘤，其特点是未分化的髓系细胞增殖不受控制，生存率低，特别是在老年患者中。AML给NHS带来了巨大的临床负担，因为控制疾病所必需的强化化疗方案往往因副作用而住院。因此，临床明显需要新的治疗方法，更具体地针对肿瘤，毒性最小。疾病复发也是AML中常见的问题，因为定义该疾病的大部分恶性未分化髓系细胞来自一种罕见的祖细胞群，称为白血病干细胞（LSC），并由其维持。LSCs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使它们对一线治疗具有抵抗力。因此，特异性去除LSCs将是AML治疗中的一个明显优势，并且有可能使用新型双特异性抗体药物偶联物（adc）来实现，这种药物偶联物能够通过结合确定的表面抗原和通过毒性物质的内化来识别和杀死这些细胞。目的：我们的主要目标是去除产生恶性髓系细胞的白血病干细胞（LSC）。我们计划与BiVictriX合作，专注于鉴定、开发和测试针对AML中LSCs的新型抗体-药物偶联物（adc）。实验方法。我们的实验方法将利用最先进的技术，涵盖免疫学、细胞生物学、分子生物学、生物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bbb40表型属性，提供基础数据，有助于加速识别新的抗原组合，为Bivictrix开发ADC靶点提供信息。来自AML和其他患者的细胞群的比较将允许识别表面抗原表型，例如但不限于表达CD34， CD38， CD133和CD90，将LSCs与非恶性对偶细胞区分开来。我们还打算通过以下组合来功能表征这些新型ADC的功效：(i)先进的显微镜技术来研究ADC在细胞系模型中的摄取和加工；（ii）一系列细胞毒性试验，在表达适当抗原的细胞系模型中测试adc的特异性和选择性；（iii）最后确定在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.