Mammary Gland as a Sensitive End Point to Effects of Endocrine Disruptors

乳腺作为内分泌干扰物影响的敏感终点

• 批准号: 10259348
• 负责人: Suzanne Fenton
• 金额: $ 340.29万
• 依托单位: NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10259348
• 关键词: 3T3-L1 Cells; Acids; Adipocytes; Adolescence; Adult; Adverse effects; Affect; Alkanesulfonates; Animal Model; Area; Arsenic; Atlases; Atrazine; Biological Assay; Birth; Breast; Breast Cancer Risk Factor; Breast Diseases; COVID-19; Carcinogens; Cells; Centers for Disease Control and Prevention (U.S.); Chemical Actions; Chemical Exposure; Chemicals; Chronic Disease; Collection; Contractor; Contracts; Core Facility; Data; Data Set; Decision Making; Development; Dose; Endocrine; Endocrine Disruptors; Endocrine Reproductive Tissue; Endocrine disruption; Environmental Health; Epidemic; Epidemiologist; Evaluation; Exposure to; Female; Fetal Development; Flame Retardants; Follow-Up Studies; Future; Gene Expression; Generations; Genes; Gland; Goals; Growth; Guidelines; Health; Hormones; Human; Hyperplasia; Impairment; In Vitro; Instruction; Interruption; Investigation; Laboratories; Lactation; Lesion; Life; Link; Liver; Malignant Neoplasms; Malignant neoplasm of male breast; Mammary Gland Parenchyma; Mammary Neoplasms; Mammary gland; Manuscripts; Melissa; Metabolism; Methodology; Military Personnel; Molecular; Mus; National Institute of Environmental Health Sciences; Obesity; Outcome; Outcome Study; Paper; Pathway interactions; Placenta; Population; Predisposition; Pregnancy; Process; Protocols documentation; Puberty; Publications; Publishing; Rattus; Regulation; Reporting; Reproductive Endocrinology; Research; Research Personnel; Risk; Risk Assessment; Robin bird; Rodent; Role; Sampling; Science; Serum; Services; Signal Transduction; Sprague-Dawley Rats; Statistical Data Interpretation; Steroids; Testing; Time; Tissues; Training; Universities; Validation; Water Supply; Weight Gain; Woman; Work; age related; analog; base; bisphenol A; blind; breast cancer diagnosis; breast density; density; developmental disease; dosimetry; environmental chemical; exposure route; girls; graduate student; human disease; improved; in silico; in vitro Assay; in vivo; interest; lipid biosynthesis; male; malignant breast neoplasm; mammary epithelium; mammary gland development; men; normal aging; novel; obesogen; obesogenic; offspring; perfluorooctanoic acid; pregnant; prenatal; prenatal exposure; pup; reproductive; response; tetrabromobisphenol A; tumor; volatile organic compound

Our research is focused on understanding the effects of environmental chemicals on mammary gland (MG) development, function, and cancer susceptibility. We have expanded this focus area in recent years to include the effect of environmental chemicals on breast cancer risk factors such as pregnancy-related birth outcomes, puberty timing, obesity, breast density and lactation. The use of animal models for human disease allows us to evaluate different routes of exposure, internal dose of the test chemical, as well as a variety of effects in the rodent that are relevant to human health. During the last several years, we have produced numerous important documents that are being used in the risk regulation of brominated flame retardants (DE-71), perfluorooctanoic acid (PFOA) and atrazine (ATR). This year, we have added GenX, a replacement chemical for PFOA, to that list. Flame retardants and perfluorinated chemicals, such as PFOA and GenX, are currently regulated at the state level, as a federal risk assessment has not been completed yet. In the female rodent offspring, the mammary gland is one of the most endocrine-sensitive end points that we have evaluated following prenatal chemical exposures. Our current and recent studies using volatile organic chemicals (VOCs), bisphenol analogues, PFOA, GenX, tetrabromobisphenol A (TBBPA), and arsenic all demonstrate this finding. We have conducted studies evaluating mammary effects of arsenic in mice, human relevant VOC mixtures in rats, PFOA and GenX mechanism of action and strain differences in mice, and two BPA replacement chemicals in mice. We have finalized our evaluation and analyses of the mammary samples from the BPA-CLARITY study and submitted the paper for publication this year. We have published the work on bisphenol analogues and mammary lesions in Environmental Health Perspectives recently and we have also recently published a manuscript comparing health effects of PFOA and GenX in pregnant mice and their developing fetuses. Numerous other manuscripts are expected from these studies in the coming year. We have made major progress on our work involving investigation of VOCs and their developmental exposure effects on the mammary gland, specifically in males. These compounds are theorized to have a role in male breast cancer diagnoses in men who were born or grew up at the Camp Lejeune military base in NC, USA. We have nearly finished dosimetry studies in the pregnant and lactating rat and her offspring in conjunction with our CDC collaborators. We have evaluated mammary gland development and carcinogen-induced mammary tumors following prenatal exposure and are finalizing manuscripts on the effects of these compounds in male and female rats. With this study and our mammary gland atlas of Harlan Sprague Dawley rats that was published two years ago, we have added significantly to what is known about male rodent mammary gland development, susceptibility to endocrine disruption, and carcinogen-induced mammary cancer. We are working with Melissa Troester, an epidemiologist at UNC, to document normal aging of the male and female mammary gland of mice and rats, in comparison to her human samples. She has shown that in women with breast cancer there is an interruption of the normal aging process, and by using chemically-exposed rodent tissues, we may determine which types of chemicals may predispose populations to increased breast cancer risk via that mode of action. Due to COVID-19, this work was not finished this year, as we had planned. In late 2018, we published a study describing susceptibility to mammary gland hyperplasia in female mice exposed to bisphenol A (BPA), BPAF and BPS (fluorinated and sulfonated forms) in early life. These studies cover important susceptible tissues not evaluated in other contracted NTP studies and add to information garnered in the BPA-CLARITY study. Dispositon studies are in their final stages to understand the transfer of these compounds to the developing fetus and the clearance within the pregnant dam and will be put into clearance soon. These studies have been delayed due to low access to mass spec time in the NIEHS core facility and in NTPL. Our recent studies involving TBBPA and Firemaster are large collaborative studies where we either conducted the study for many PIs to get target tissues of interest (TBBPA) or we are providing a service of isolating and evaluating mammary tissue (Firemaster). Two papers were published in this reporting year and three remain, but have been delayed due to COVID-19 and access to the lab. Both of these flame retardants seem to act as endocrine disruptors, affecting multiple reproductive tissues, fat cells, and hormone levels. We are finalizing the work on a blind evaluation of about 70 chemicals identified in Tox21 as activating specific endpoints thought to be linked to an obesogenic response. We are using mouse 3T3-L1 low passage cells to assess adipogenesis and lipogenesis. Gene expression and follow-up studies have begun on the chemicals identified as most interesting, either for their potential to act as a potent obesogen or because they were a false positive or negative. We are working with a team of investigators, including Kris Thayer at US EPA, David Allen at ILS, Inc., and Jui-Hua Hseih in BSB,to produce a document on this concept and test other aspects of the chemical response. From the data accumulated in this group effort, we will better understand the utility and shortcomings of the Tox21 data set in predicting obesogens and may be able to develop assays to add to the high throughput methodology. We have completed this year a round-robin global validation for the 3T3-L1 cells for use in these assays, which was coordinated by Chris Kassotis, of Wayne State University. Some of the reporting was delayed due to COVID-19 and access to lab. Since 2018, there has been an emergent need to evaluate the effects of uncharacterized perfluoroalkyl acids (PFAS) and related compounds for health effects and mechanisms of action. We have developed a set of in vitro assays (complimented by our colleagues in the in silico and in vivo areas) that will provide us an our collaborators at the EPA with a rapid understanding of which groups of PFAS are targeting tissues known to be affected by the legacy compounds before them that are no longer produced in the U.S. We are testing nearly 50 of the most common compounds in liver, mammary cells, placenta, and fat cells to determine bench mark doses across the group. We have recently finished two manuscripts from this work and they are being prepared for clearance. Additionally we co-authored papers with Rebecca Fry's lab describing gene changes in human placental cells following serum plus/minus culture conditions. These important contributions will move the field forward in high throughput testing of the PFAS. We are also testing AFFF mixtures in vitro and enhancing those assays with single compounds identified in the mass spec data supplied by our EPA and Harvard collaborators. One of these papers has gone through clearance and another will be there shortly. Our data is needed by the state of NC and the Dept of Defense in the near future to make decisions on new product use, regulation or filtering of water supplies, and potentially health advisories. We will be very busy in this area in the coming years. One manuscript investigating the relationship of chronic disease in human populations with PFAS exposures was published last year, with Kelly Ferguson as the senior investigator, as she trained my graduate student in statistical analyses needed for that work. Our work comparing GenX and PFOA effects on the placenta was published recently and the studies on offspring growth and mammary development are in final form and being readied for NIEHS clearance.

我们的研究重点是了解环境化学物质对乳腺 (MG) 发育、功能和癌症易感性的影响。近年来，我们扩大了这一重点领域，纳入了环境化学物质对乳腺癌危险因素的影响，例如与妊娠相关的出生结果、青春期时机、肥胖、乳房密度和哺乳期。使用人类疾病动物模型使我们能够评估不同的暴露途径、测试化学品的内部剂量以及与人类健康相关的啮齿动物的各种影响。 在过去几年中，我们编写了许多重要文件，用于溴化阻燃剂 (DE-71)、全氟辛酸 (PFOA) 和莠去津 (ATR) 的风险监管。今年，我们将 PFOA 的替代化学品 GenX 添加到该列表中。由于联邦风险评估尚未完成，阻燃剂和全氟化学品（例如 PFOA 和 GenX）目前在州一级受到监管。 在雌性啮齿动物后代中，乳腺是我们在产前化学暴露后评估的最内分泌敏感的终点之一。我们当前和最近使用挥发性有机化学品 (VOC)、双酚类似物、PFOA、GenX、四溴双酚 A (TBBPA) 和砷的研究都证明了这一发现。我们开展的研究评估了砷对小鼠乳腺的影响、人类相关 VOC 混合物对大鼠的影响、PFOA 和 GenX 的作用机制以及小鼠品系差异，以及两种 BPA 替代化学物质对小鼠的影响。我们已经完成了对 BPA-CLARITY 研究中乳腺样本的评估和分析，并于今年提交了论文发表。我们最近在《环境健康展望》中发表了有关双酚类似物和乳腺病变的研究，最近还发表了一篇手稿，比较了 PFOA 和 GenX 对怀孕小鼠及其发育中胎儿的健康影响。预计这些研究将在来年发表许多其他手稿。 我们在研究挥发性有机化合物及其对乳腺（特别是男性）发育暴露影响的工作方面取得了重大进展。理论上，这些化合物在美国北卡罗来纳州勒琼军营军事基地出生或长大的男性乳腺癌诊断中发挥作用。我们与疾病预防控制中心的合作者合作，即将完成对怀孕和哺乳期大鼠及其后代的剂量测定研究。我们评估了产前接触后乳腺发育和致癌物诱发的乳腺肿瘤，并正在最终确定这些化合物对雄性和雌性大鼠的影响的手稿。通过这项研究和我们两年前发表的 Harlan Sprague Dawley 大鼠乳腺图谱，我们显着增加了对雄性啮齿动物乳腺发育、内分泌干扰易感性和致癌物诱发乳腺癌的了解。我们正在与北卡罗来纳大学的流行病学家 Melissa Troester 合作，记录小鼠和大鼠的雄性和雌性乳腺的正常衰老情况，并与她的人类样本进行比较。她表明，患有乳腺癌的女性的正常衰老过程会受到干扰，通过使用接触化学物质的啮齿动物组织，我们可以确定哪些类型的化学物质可能通过这种作用方式使人群易患乳腺癌的风险增加。由于 COVID-19，这项工作今年没有按我们的计划完成。 2018 年底，我们发表了一项研究，描述了生命早期接触双酚 A (BPA)、BPAF 和 BPS（氟化和磺化形式）的雌性小鼠对乳腺增生的易感性。这些研究涵盖了其他合同 NTP 研究中未评估的重要易感组织，并补充了 BPA-CLARITY 研究中收集的信息。处置研究正处于最后阶段，以了解这些化合物向发育中的胎儿的转移以及怀孕母体内的清除，并将很快投入清除。由于 NIEHS 核心设施和 NTPL 获取质谱的时间较少，这些研究已被推迟。我们最近涉及 TBBPA 和 Firemaster 的研究是大型合作研究，我们要么为许多 PI 进行研究以获得感兴趣的目标组织 (TBBPA)，要么提供分离和评估乳腺组织的服务 (Firemaster)。本报告年度发表了两篇论文，还有三篇论文发表，但由于 COVID-19 和进入实验室的原因而被推迟。这两种阻燃剂似乎都会起到内分泌干扰物的作用，影响多种生殖组织、脂肪细胞和激素水平。 我们正在完成对 Tox21 中确定的约 70 种化学物质的盲评估工作，这些化学物质可激活被认为与肥胖反应相关的特定终点。 我们使用小鼠 3T3-L1 低传代细胞来评估脂肪生成和脂肪生成。对被认为是最有趣的化学物质的基因表达和后续研究已经开始，要么是因为它们具有作为有效致肥胖剂的潜力，要么是因为它们是假阳性或阴性。我们正在与一个研究团队合作，包括美国 EPA 的 Kris Thayer、ILS, Inc. 的 David Allen 和 BSB 的 Jui-Hua Hseih，编写有关这一概念的文件并测试化学反应的其他方面。从该小组工作中积累的数据中，我们将更好地了解 Tox21 数据集在预测肥胖因素方面的效用和缺点，并可能开发出检测方法以添加到高通量方法中。我们今年完成了对用于这些测定的 3T3-L1 细胞的循环全球验证，该验证由韦恩州立大学的 Chris Kassotis 协调。由于 COVID-19 和进入实验室的原因，一些报告被延迟。 自 2018 年以来，迫切需要评估未表征的全氟烷基酸 (PFAS) 和相关化合物对健康的影响和作用机制。 我们开发了一套体外测定（得到了我们在计算机和体内领域的同事的称赞），这将使我们在 EPA 的合作者能够快速了解​​哪些 PFAS 组针对已知受美国不再生产的遗留化合物影响的组织。我们正在测试肝脏、乳腺细胞、胎盘和脂肪细胞中近 50 种最常见的化合物，以确定 整个组的基准剂量。我们最近完成了这项工作的两份手稿，正在准备审批。 此外，我们与 Rebecca Fry 的实验室共同撰写了论文，描述了在血清加/减培养条件下人类胎盘细胞的基因变化。这些重要贡献将推动 PFAS 高通量测试领域的发展。我们还在体外测试 AFFF 混合物，并使用我们的 EPA 和哈佛大学合作者提供的质谱数据中鉴定的单一化合物来增强这些测定。其中一份文件已通过审批，另一份文件很快就会到达。北卡罗来纳州和国防部在不久的将来需要我们的数据来就新产品的使用、供水的监管或过滤以及潜在的健康建议做出决策。未来几年我们将在这一领域非常忙碌。去年发表了一份调查人类慢性疾病与 PFAS 暴露之间关系的手稿，凯莉·弗格森 (Kelly Ferguson) 担任高级研究员，她对我的研究生进行了这项工作所需的统计分析方面的培训。我们最近发表了比较 GenX 和 PFOA 对胎盘影响的研究成果，有关后代生长和乳腺发育的研究已形成最终版本，正在为 NIEHS 审批做好准备。