Sensitizing Leukemia Stem Cells to NF-kB Inhibition by Blocking TNF Signaling

通过阻断 TNF 信号传导使白血病干细胞对 NF-kB 抑制敏感

• 批准号: 8456470
• 负责人: Andrew Volk
• 金额: $ 2.92万
• 依托单位: LOYOLA UNIVERSITY CHICAGO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-03-08 至 2016-03-07
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8456470
• 关键词: Acute Myelocytic Leukemia; Adverse effects; American; Apoptosis; Biological Assay; Blood Cells; Bone Marrow; Cell Death; Cell Proliferation; Cell Survival; Cell physiology; Cells; Cessation of life; Clinic; Clinical; Data; Development; Disease; Dose; Elements; Equilibrium; Future; Genes; Genetic; Goals; Hematopoietic; Hematopoietic stem cells; Homeostasis; Human; Immune system; In Vitro; Inflammation; Inflammatory; Integumentary System Part; Knock-out; Leukemic Cell; Liver; MAP3K7 gene; MAPK14 gene; MAPK8 gene; MLL-AF9; Measures; Mediating; Modeling; Monoclonal Antibodies; Mus; NF-kappa B; Nervous system structure; Newly Diagnosed; Pancytopenia; Pathogenesis; Pathway interactions; Patients; Phosphotransferases; Proto-Oncogene Proteins c-akt; Receptor Activation; Receptor Signaling; Relapse; Role; Sampling; Signal Pathway; Signal Transduction; Skin; Staining method; Stains; Stem cells; Testing; Therapeutic; Tissues; Toxic effect; Transplantation; Tumor Necrosis Factor Receptor; Tumor Necrosis Factor-alpha; annexin A5; caspase-8; cell growth; cytokine; enhancing factor; fusion gene; hematopoietic tissue; human TNFRSF1A protein; in vivo; inhibitor/antagonist; killings; leukemia; leukemic stem cell; novel; novel strategies; novel therapeutics; prevent; progenitor; public health relevance; receptor; research study; small molecule; treatment effect; tumor necrosis factor alpha receptor

DESCRIPTION (provided by applicant): Leukemia stem cells (LSCs) are the key subset of leukemic cells (LCs) required for leukemia initiation, progression and relapse. Complete elimination of LSCs is currently the ultimate goal of leukemia therapy. In 2001, it was found that using an inhibitor against NF-?B (a transcriptional regulator of many survival genes) resulted in significant death of LSCs in human acute myeloid leukemia samples while preserving healthy hematopoietic stem cells /progenitors (HSC/Ps).1 Unfortunately, the clinical use of NF-?B inhibitors at therapeutic levels is limited by the development of severe inflammation-related side-effects in the skin, the liver, and elements of the nervous, and immune systems. These side effects were uncovered during testing in mice.2-4. Therefore, we need new therapies that reduce side-effects either by using a better NF-?B inhibitor at a lower dose or by developing novel therapies which prevent the inflammation caused by NF-?B inhibitor. Tumor Necrosis Factor alpha (TNF) is a major inflammatory signal released by hematopoietic cells and is implicated in NF-?B modulation therapies.5, 6 While TNF's inflammatory signals causes cell death and growth arrest in normal HSC/Ps, its role in leukemia development is not yet clear.7, 8 What is clear is that managing inflammation is critical for the successful treatment of leukemia.9, 10 Our preliminary studies show that LCs utilize TNF-mediated inflammation as both a survival and proliferation signal by activating a signaling network that depends in part on NF-?B. By knocking out both receptors of TNF (Tnfr1a-/-1b-/-), we can sensitize LCs 100-fold to NF-?B inhibitor treatment when compared to normal HSC/Ps. Therefore, we hypothesize that inhibiting both TNF receptor signaling and NF-?B would provide a double benefit: clinicians could use NF-?B inhibitors at a lower dose while simultaneously blocking TNF-dependent inflammation. Our broad goal in this study is to identify novel leukemia-specific therapeutic strategies that can augment NF-?B inhibition. Our immediate goal is to determine whether blocking TNF signaling augments NF- ?B inhibition in eliminating both leukemic cells and leukemic stem cells. In our study, we will compare treatments in LCs and bone marrow (BM) HSC/Ps, and will take advantage of the murine genetic leukemia models we have generated. We will utilize pharmacological approaches to alter the activity of signaling pathways in order to maximally kill LCs while preserving normal HSC/Ps. We will measure global effects on cell death by Annexin V/PI staining, colonigenic capacity of progenitor cells by CFU assay, and hematopoietic reconstitutive capacity of normal HSC/Ps and leukemogenic capacity of LSCs by in vivo transplantation. The first aim of our study explores the effects of blocking TNF in conjunction with NF-?B inhibitor treatment. Our second aim explores small molecule inhibition of TNF receptor downstream targets in combination with NF-?B inhibitor treatment. The expected results of this study will significantly contribute to furthering the understanding of leukemia, an may also point the way toward novel strategies for future therapies.

描述（由申请人提供）：白血病干细胞（LSC）是白血病启动，进展和复发所需的白血病细胞（LCS）的关键子集。目前，完全消除LSC是白血病疗法的最终目标。在2001年，发现使用针对NF-？B（许多生存基因的转录调节剂）的抑制剂会导致人类急性髓性白血病样本中LSC的显着死亡，同时保留健康的造血干细胞 /祖细胞 /祖细胞 /祖细胞 /祖细胞 /祖细胞（HSC /PS）。不幸的是，通过临床限制了在NF-的临床水平上，在临床上的使用量有限，在更稳定的水平上，在抑制型抑制剂中均抑制了抑制剂，该抑制剂均为抑制型抑制剂。神经和免疫系统的皮肤，肝脏和元素中的副作用。这些副作用在小鼠测试过程中被发现。2-4。因此，我们需要通过在较低剂量下使用更好的NF-？B抑制剂或开发新的疗法来减少副作用的新疗法，从而可以防止NF-抑制剂引起的炎症。 肿瘤坏死因子α（TNF）是造血细胞释放的主要炎症信号，与NF-？B调节疗法有关。5，6，而TNF的炎症信号在正常的HSC/PS中导致细胞死亡和生长在正常的HSC/PS中导致其在白血病中的作用，其在白血病中的作用尚不清楚。7尚未清楚地处理。7，8是什么，这是对液压的关键。 10我们的初步研究表明，LCS利用TNF介导的炎症作为生存和增殖信号，通过激活部分取决于NF-？b。通过淘汰TNF的两个受体（TNFR1A - / - 1B - / - ），与正常的HSC/PS相比，我们可以将LCS 100倍对NF-？B抑制剂治疗敏感。因此，我们假设抑制TNF受体信号传导和NF-？b将提供双重益处：临床医生可以以较低剂量使用NF-？B抑制剂，同时抑制TNF依赖性炎症。 我们在这项研究中的广泛目标是确定可以使用新颖的白血病特异性治疗策略 增强NF-？B抑制作用。我们的近期目标是确定阻断TNF信号传导是否会增加NF-？b消除白血病细胞和白血病干细胞的抑制作用。在我们的研究中，我们将比较LCS和骨髓（BM）HSC/PS的治疗方法，并将利用我们生成的鼠遗传性白血病模型。我们将利用药理方法改变信号通路的活性，以最大程度地杀死LC，同时保留正常的HSC/PS。我们将通过Annexin V/PI染色，CFU测定法对祖细胞的结肠融合能力以及正常HSC/PS的造血重建能力以及LSC通过体内移植的LSC的白血病能力来衡量全球对细胞死亡的影响。我们研究的第一个目的探讨了阻断TNF与NF-抑制剂治疗结合的影响。我们的第二个目标探索了TNF受体下游靶标的小分子抑制，并结合NF-抑制剂治疗。这项研究的预期结果将极大地有助于进一步了解白血病的理解，这也可能指向迈向未来疗法的新型策略的道路。