EARLY DETECTION OF CLONAL HEMATOPOIESIS AND LEUKEMIA ASSOCIATED MUTATIONS IN WTC EXPOSED FIREFIGHTERS AFTER THE 9/11 ATTACKS

摘要 世界贸易中心（WTC）的恐怖袭击造成了前所未有的环境暴露， WTC雾化的灰尘和气体含有已知和可疑的致癌物质，包括多环芳烃 碳氢化合物、多氯联苯、呋喃、二恶英和石棉。我们小组的研究报告说 暴露于WTC的纽约市消防局（FDNY）消防员中的癌症病例过多，（2， 3)包括白血病和多发性骨髓瘤发病率上升的趋势。我们继续前瞻性地 跟踪整个纽约消防局消防员队列的癌症发病率，并建立了一个样本生物储存库 由2000多名暴露于WTC的消防员和非WTC消防员的分馏血液成分和血清组成， 暴露的消防员控制装置使用这个队列，我们对781名WTC暴露者进行了初步研究。 消防队员表现出显着较高的单克隆丙种球蛋白病的患病率， 重要性（MGUS），多发性骨髓瘤的前兆。(4)此外，我们还进行了深入的针对性研究。 暴露于WTC的消防员（N=481）和年龄匹配的未暴露于WTC的消防员的试点队列的基因组测序 消防员对照组（N=52）。我们观察到克隆性造血（CH）和白血病相关的高发病率， 突变（57/481; 12%）在WTC暴露的消防员。对照组的突变率为2%， 与已发表研究中的年龄匹配对照组相当（5，6）。克隆性造血（CH）是获得 白血病相关的体细胞突变增加了血液癌和所有原因的风险， mortality.在暴露于WTC的消防员中观察到的突变影响了DNMT 3A，TET 2，TP 53，剪接和其他基因。 通常是反复突变的基因。这些突变表明CH的发生率很高，而其他队列中的CH 已被证明与骨髓肿瘤、炎症和心脏病的发生风险有关。 疾病我们现在建议全面确定CH的患病率在一个大型FDNY WTC暴露 队列和研究由WTC颗粒物（WTC- PM）。目的1将使用深度靶向测序确定克隆性造血突变的患病率， 暴露于WTC的消防员的大队列和对照样品的2个队列（非WTC消防员和非WTC消防员）。 消防员年龄匹配对照）。突变将与临床变量相关，如WTC水平 暴露、年龄、连续血细胞计数、吸烟史和其他临床特征。目标2将决定 WTC颗粒物对小鼠体内克隆性造血发育的功能影响 TET 2和DNMT 3A基因突变的模型。最后，目标3将决定WTC-PM的效果 暴露于单分子水平的细胞变化，以评估DNA损伤的机制和程度 暴露细胞的损伤。这些结果将进一步加深我们对该机制的理解， 环境暴露导致基因组不稳定。重要的是，CH突变的早期检测将使 对暴露于WTC的消防员进行有针对性的临床评估，以便早期诊断和改变潜在的疾病 干预措施-NIOSH WTC健康计划的“研究到护理”转化模式的中心目标。

ABSTRACT The terrorist attacks on the World Trade Center (WTC) created an unprecedented environmental exposure to WTC aerosolized dust and gases that contained known and suspected carcinogens including polycyclic aromatic hydrocarbons, polychlorinated biphenyls, furans, dioxins and asbestos. Studies from our group have reported an excess of cancer cases in the WTC-exposed Fire Department of the City of New York (FDNY) firefighters,(2, 3) including a trend towards higher incidence of leukemia and multiple myeloma. We continue to prospectively follow the entire FDNY cohort of firefighters for cancer incidence and have built a biorepository of samples consisting of fractionated blood components and serum from 2000+ WTC-exposed firefighters and non-WTC- exposed firefighter controls. Using this cohort, we have conducted preliminary studies in 781 WTC-exposed firefighters that demonstrate a significantly higher prevalence of monoclonal gammopathy of undetermined significance (MGUS), a precursor for multiple myeloma. (4) Furthermore, we have conducted deep targeted genome sequencing of a pilot cohort of WTC-exposed firefighters (N=481) and non-WTC-exposed age matched firefighter controls (N=52). We observed a high incidence of Clonal hematopoiesis (CH) and leukemia associated mutations (57/481; 12%) in WTC-exposed firefighters. The rate of mutations in controls was 2% and is comparable to age matched controls in published studies (5, 6). Clonal hematopoiesis (CH) is acquisition of leukemia associated somatic mutations that carry increase risk of hematologic cancer as well as all-cause mortality. The mutations seen in WTC-exposed firefighters affected DNMT3A, TET2, TP53, Splicing and other commonly recurrently mutated genes. These mutations suggest a high rate of CH, and CH in other cohorts has been shown to be associated with risk for development of myeloid neoplasms, inflammatory conditions and heart diseases. We now propose to comprehensively determine the prevalence of CH in a large FDNY WTC-exposed cohort and study the molecular and cellular mechanisms of damage caused by WTC particulate matter (WTC- PM). Aim 1 will determine the prevalence of clonal hematopoiesis mutations using deep targeted sequencing in a large cohort of WTC-exposed firefighter and 2 cohorts of control samples (non-WTC firefighters and non- firefighter age matched controls). Mutations will be correlated with clinical variables such as level of WTC exposure, age, serial blood counts, smoking history and other clinical characteristics. Aim 2 will determine the functional effects of WTC particulate matter on development of clonal hematopoiesis in vivo by using mouse models with commonly mutated TET2 and DNMT3A genes. Lastly, aim 3 will determine the effect of WTC-PM exposure on cellular changes at the single molecule level to assess the mechanisms and magnitude of DNA damage in exposed cells. These results will further our understanding on the mechanism via which environmental exposures lead to genomic instability. Importantly, early detection of CH mutations will enable targeted clinical evaluations of WTC-exposed firefighters, enabling early diagnosis and potential disease altering interventions – a central goal of the NIOSH WTC Health Program's “Research to Care” translational model..