Stimulated Differentiation of Native Human Acute Myeloid Leukemia (AML) Blasts as a Potential Cancer-Cell Specific Therapeutic Target - Part 1, 2010

The survey "Stimulated Differentiation of Native Human Acute Myeloid Leukemia (AML) Blasts as a Potential Cancer-Cell Specific Therapeutic Target, 2010" consists of three parts. These are:

Del 1: NSD1474-1 - Reticulated plates with hematologic reconstitution of the bone marrow after chemotherapy.
Del 2: NSD1474-2 - Differentiating the treatment of acute myeloid leukemia.
Del 3: NSD1474-3 - Autophagy.

Acute myeloid leukemia (AML) is an aggressive disorder characterized by accumulation in the bone marrow of large numbers of abnormal blood cells that fail to develop into normal functional blood cells. This is a life-threatening condition if not treated. Patients under the age of 60 are given intensive cytotoxic treatment, but the five year survival after diagnosis of AML is below 20 %. Most patients diagnosed with AML are above 60 years, and there is a need for newer and more effective and targeted treatments with acceptable side. The present project primarily aims at stimulating the halted maturation of native human AML progenitors. Clinically, differentiation-stimulating treatment with a combination of all-trans retinoicacid (ATRA)+valproat+theophyllamine will be given to patients and the expression of differentiation associated membrane molecules and phosphorylation status of intracellular signal transduction pathways will be characterized. The effect of this treatment will be characterized in detail both on leukemic and normal bloodcells. Experimental studies will elucidate further the degree of apoptosis, proliferation, cell cycle and differentiation (morphologic and certain immunologic markers), known signal transduction pathways (protein- and lipid phosphorylation, tumor suppressor gene products. The balance between the two gene-expression regulating enzymes histone deacetylase (HDAC) and DNA methyltransferase (DMT) by inhibitors is a potential target for treatment of cancer cells and the combination of HDAC and DMT inhibitors (with or without growth factors) is very effective in inducing differentiation, apoptosis, and/or growth arrest in leukemia. We will investigate the effect of these inhibitors on differentiation for a large group of native AML blasts and xenograft models (NOD/SCID) with regard to proliferation, apoptosis, markers for normal hematopoietic progenitor differentiation and cellular signaling processes that might be affected.

The prosjects main goal and its learning objective:
The main goal is to induce differentiation of immature non-functional native human AML blasts (clinically and experimentally), and thereby produce regular cell cycles and expression of tumor suppressor gene products as a strategy to achieve normal hematopoietic maturation. Clinical studies with combination-treatment (all-trans retinoicacid (ATRA)+valproat+theophyllamine) on expression of differentiation associated membrane molecules and phosphorylation status of intracellular signal transduction pathways. Experimental studies will elucidate further the degree of apoptosis, proliferation, cell cycle and differentiation (morphologic and certain immunologic markers), known signal transduction pathways (protein- and lipid phosphorylation and tumor suppressor gene products. AML cell populations with or without differentiation-therapy will be applied to xenograft models with NOD/SCID mouse, engraftment and the degree of leukemia development as well as optical imaging of engraftment

调查《2010年：天然人类急性髓系白血病（AML）原始细胞的诱导分化作为潜在的癌细胞特异性治疗靶点》包含三个部分，具体如下：

部分1（NSD1474-1）：化疗后骨髓造血重建相关的网状板
部分2（NSD1474-2）：急性髓系白血病（AML）治疗的差异化研究
部分3（NSD1474-3）：自噬

急性髓系白血病（AML）是一种侵袭性疾病，其特征为骨髓中大量异常血细胞积聚，这些细胞无法发育为正常功能的血细胞。若不及时治疗，该病会危及生命。60岁以下患者接受强化细胞毒性治疗，但AML确诊后的5年生存率仍低于20%。大多数AML患者年龄超过60岁，因此亟需更有效、更具针对性且副作用可控的新型治疗方案。本项目主要旨在刺激天然人类AML祖细胞停滞的成熟过程。临床上，将采用全反式维甲酸（ATRA）+丙戊酸盐+茶碱的联合分化诱导治疗方案，对患者进行干预，并表征分化相关膜分子的表达及细胞内信号转导通路的磷酸化状态。该治疗对白血病细胞及正常血细胞的影响将被详细表征。实验研究将进一步阐明凋亡、增殖、细胞周期及分化（形态学及特定免疫学标志物）的程度，以及已知的信号转导通路（蛋白质与脂质磷酸化、抑癌基因产物）。组蛋白脱乙酰酶（HDAC）与DNA甲基转移酶（DMT）这两种基因表达调控酶的抑制剂平衡，是癌细胞治疗的潜在靶点；HDAC与DMT抑制剂（联合或不联合生长因子）在诱导白血病细胞分化、凋亡及/或生长停滞方面极为有效。我们将针对大量天然AML原始细胞及异种移植模型（NOD/SCID），探究这些抑制剂对分化的影响，涉及增殖、凋亡、正常造血祖细胞分化标志物及可能受影响的细胞信号过程。

项目的主要目标及学习目标：
主要目标是（通过临床与实验手段）诱导未成熟、无功能的天然人类AML原始细胞分化，进而产生规律的细胞周期并表达抑癌基因产物，以此作为实现正常造血成熟的策略。开展联合治疗（全反式维甲酸（ATRA）+丙戊酸盐+茶碱）的临床研究，分析分化相关膜分子的表达及细胞内信号转导通路的磷酸化状态。实验研究将进一步阐明凋亡、增殖、细胞周期及分化（形态学及特定免疫学标志物）的程度，以及已知的信号转导通路（蛋白质与脂质磷酸化、抑癌基因产物）。将接受或未接受分化治疗的AML细胞群应用于NOD/SCID小鼠异种移植模型，观察移植成功率、白血病发展程度及移植的光学成像情况。