Time to ATTAC: Adoptive Transfer of T cells Against gp100+ Cells to treat LAM

摘要 淋巴管肌瘤病(LAM)是一种罕见的疾病，对年轻女性造成毁灭性的后果。 被诊断出患有这种疾病。我们观察到糖蛋白gp100和其他黑色素瘤的隐蔽表达。 结节状、肺部肿瘤病变中的相关抗原。LAM细胞携带功能失调的TSC复合体， 表现出结构性的mTOR激活。虽然雷帕霉素可以提供缓解，但非常需要开发一种 真正治愈患有LAM的女性。我们评估了合适的抗原的表达和相关的渗透。 通过免疫细胞。在这里，我们建议利用黑色素瘤相关抗原的隐蔽表达来 为LAM开发安全有效的T细胞免疫疗法。首先，我们将生成目标构造 LAM抗原转导T细胞，并为过继转移准备细胞。这些构造配备或 不使用归巢受体来驱动过继转移的T细胞到LAM肺，利用一致的 在受影响组织中过度表达趋化因子配体CCL2，以最大限度地减少对肿瘤的影响。 转基因T细胞的产生具有干细胞样的属性，以促进寿命和持续功能。 TCR转基因和非人类白细胞抗原非依赖性CAR T细胞靶向的有效性和安全性比较 A LAM表面抗原。基于构建体编码的天然受体表位的表达，结果T 细胞很容易被分类和追踪，并将在疾病的小鼠模型中进行测试。我们将与 一种免疫活性疾病模型，以及PDX移植小鼠，以探索治疗潜力 过继转移的LAM反应性T细胞。在PDX小鼠体内，LAM微环境被很好地保存。 LAM肿瘤抗原的表达可以随着时间的推移而保持，而治疗方法可以在统计上进行测试 在生物学上有意义的方式。最后，我们将探讨LAM和PDX的免疫微环境。 组织是否接受过继转移，以了解T细胞是由LAM肺容纳还是由LAM提供 可以教会采用转移术来清除现有的病变。下一步，我们的协作小组旨在开发一个双赢的 免疫治疗方法来治疗这种毁灭性的疾病。因此，该项目将包括(1)发电和在 转基因小鼠和人类T细胞的体外检测；以及(2)过继抗肿瘤效果的测量 LAM免疫活性和PDX模型中转移的T细胞以及(1)深入分析 过继转移前后LAM皮损中的免疫环境。由此产生的临床前数据 然后可以用于在后续研究中设计临床试验并检验假设，即LAM中的良性肿瘤 可通过组织归巢、LAM反应性T细胞过继转移治疗。

SUMMARY Lymphangioleiomyomatosis (LAM) is a rare disorder with devastating consequences for the young women diagnosed with this disease. We observed occult expression of the glycoprotein gp100 and other melanoma associated antigens in nodular, pulmonary tumor lesions. LAM cells carry a dysfunctional TSC complex and exhibit constitutive mTOR activation. Though rapamycin can provide relief, there is a great need to develop a true cure for women with LAM. We evaluated the expression of suitable antigens and the associated infiltration by immune cells. Here we propose to capitalize on the occult expression of melanoma associated antigens to develop safe and effective, T cell-based immunotherapy for LAM. First, we will generate constructs targeting LAM antigens to transduce T cells and prepare the cells for adoptive transfer. These constructs are equipped or not with a homing receptor to drive the adoptively transferred T cells to LAM lung, exploiting the consistent overexpression of the chemokine ligand CCL2 within affected tissues., in order to minimize off tumor effects. Transgenic T cells are generated with stem cell-like attributes to promote longevity and continued functionality. Efficacy and safety comparisons are made between TCR transgenic and HLA-independent CAR T cells targeting a LAM surface antigen. Based on expression of a natural receptor epitope encoded by the construct, resulting T cells are readily sorted and traced, and will be put to the test in mouse models of the disease. We will engage an immunocompetent disease model, as well as PDX implanted mice to explore the treatment potential of adoptively transferred, LAM reactive T cells. Within PDX mice, the LAM microenvironment is well conserved. Expression of LAM tumor antigens can be maintained over time, while therapeutics can be tested in a statistically and biologically meaningful way. Finally, we will explore the immune microenvironment in LAM and in PDX tissues exposed to adoptive transfer or not, to learn whether T cells harbored by LAM lung or supplied by adoptive transfer can be taught to clear existing lesions. Next, our collaborative group aims to develop a winning immunotherapeutic approach to treat the devastating disease. The project will thus cover (1) generating and in vitro testing of transgenic mouse and human T cells; and (2) measuring the anti-tumor efficacy of adoptively transferred T cells in immune competent as well as PDX models of LAM as well as (1) in-depth analysis of the immune environment encountered in LAM lesions before and after adoptive transfer. Resulting preclinical data can then serve to design a clinical trial in follow-up studies and test the hypothesis, that benign tumors in LAM are amenable to treatment by adoptive transfer of tissue homing, LAM reactive T cells.