Sustained regulation of hypothalamus-pituitary-ovary hormones with tissue-engineered ovarian constructs as a treatment for osteoporosis in females

利用组织工程卵巢结构持续调节下丘脑-垂体-卵巢激素作为女性骨质疏松症的治疗方法

• 批准号: 10659277
• 负责人: Barbara D. Boyan
• 金额: $ 51.8万
• 依托单位: MIAMI UNIVERSITY OXFORD
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-15 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10659277
• 关键词: Address; Affect; Age; Age Years; Alendronate; Alginates; Allogenic; American; Animals; Anterior Pituitary Gland; Belief; Biomimetics; Cardiovascular Diseases; Cardiovascular system; Cell Separation; Cell Survival; Cell Therapy; Cells; Clinic; Clinical; Development; Direct Costs; Dose; Encapsulated; Endocrine; Enzyme-Linked Immunosorbent Assay; Epigenetic Process; Estrogens; Female; Follicle Stimulating Hormone; Fracture; Glucocorticoids; Goals; Heart; Histology; Hormone secretion; Hormones; Human; Hypothalamic Hormones; Hypothalamic structure; Immune response; Immunologics; Implant; In Vitro; Incidence; Knowledge; Luteinizing Hormone; Malignant Neoplasms; Measures; Mechanics; Menopause; Messenger RNA; MicroRNAs; Modeling; Myocardial Infarction; Oocytes; Oryctolagus cuniculus; Osteoporosis; Osteoporosis prevention; Outcome; Ovarian; Ovarian Follicle; Ovarian Granulosa Cell; Ovarian Tissue; Ovarian hormone; Ovariectomy; Ovary; Pathway interactions; Performance; Pharmaceutical Preparations; Pharmacological Treatment; Physiological; Pituitary Gland; Pituitary Hormones; Plasma; Postmenopause; Premature Ovarian Failure; Prevalence; Progesterone; Rattus; Recommendation; Regulation; Regulatory Pathway; Research; Risk; Risk Reduction; Safety; Source; Stroke; Structure; Surgical Models; System; Testing; Time; Tissue Engineering; Tissue-Specific Gene Expression; Tissues; Uterus; Woman; Women' s Health; bisphosphonate; bone; bone health; cancer type; cell type; clinical translation; confocal imaging; design; egg; experimental study; gender difference; granulosa cell; hormone therapy; hospitalization rates; implantation; improved; in silico; insight; loss of function; malignant breast neoplasm; men; microCT; next generation sequencing; older women; osteoporosis with pathological fracture; osteoporotic bone; ovarian failure; pharmacologic; polyornithine; safety outcomes; theca cell; therapeutic target; transcriptome sequencing

SUSTAINED REGULATION OF HYPOTHALAMUS-PITUITARY-OVARY HORMONES WITH TISSUE- ENGINEERED OVARIAN CONSTRUCTS AS A TREATMENT FOR OSTEOPOROSIS IN FEMALES PROJECT SUMMARY/ABSTRACT Females are disproportionately affected by osteoporosis and osteoporotic fracture. In the U.S., approximately 40 million women have or are at risk of developing osteoporosis and the majority (~ 60%) of the 2 million osteoporotic bone fractures (direct costs > $20 billion per year) occur in women. Women have a 1-in-2 lifetime chance of having an osteoporotic fracture and a 4-fold higher rate of osteoporosis than men. A key underlying cause is ovarian failure due to menopause or other conditions that lead to loss of ovarian hormones with concomitant disruption of the hypothalamus-ovary-pituitary (HPO) axis. Until 2002, traditional pharmacological hormone therapy (pHT) was widely used due to its perceived ability to reduce risk of osteoporosis and, importantly, osteoporotic fracture. The Women’s Health Initiative (WHI) verified this long-held belief, demonstrating that pHT reduced incidence of osteoporotic fracture. However, the WHI also indicated that the risks of hormone therapy, such as cardiovascular disease and certain types of cancer, outweighed the benefits of reduced levels of osteoporosis. Follow-on studies indicate that risks may be higher in women older than 60 years of age and/or more than 10 years post-menopause, suggesting that hormone therapy may still be effective if given at lower, safer doses and via suitable delivery platforms. We propose biomimetic, ovarian cell constructs as a tissue engineering approach to hormone delivery (cellular hormone therapy; cHT) in the treatment of osteoporosis. The hypothesis guiding this research is that cHT can achieve better bone health and safety outcomes than pharmacological agents, such as pHT or the bisphosphonates, in ovarian failure because it mimics native ovarian structure and function. Our cHT approach uses a spatial arrangement of two key ovarian cell types (granulosa and theca cells) that mimics native ovarian follicle structure while avoiding pitfalls of whole ovarian tissue encapsulation such as oocyte (egg) expulsion and loss of function with time. Our system has achieved sustained and physiologically- relevant levels of ovarian hormone secretion. We have demonstrated that the encapsulated ovarian cells participate in the HPO axis, as evidenced by regulation of follicle stimulating hormone (FSH) and luteinizing hormone (LH) levels from the anterior pituitary. There are fundamental differences between our cHT constructs and the native ovary (e.g., lack of oocytes in our constructs) but ovarian, pituitary, and possibly hypothalamic hormones are regulated in a manner similar to a pre-ovarian failure state. These cell-based constructs also release other hormones naturally secreted by native ovaries but not present in pHT. More importantly, cHT achieved beneficial bone outcomes in a manner that was safer than pHT. The long-term goal of this research is to develop a cell-based therapy that can be used in humans for the prevention of osteoporosis associated with loss of ovarian function in women. In our preliminary studies, we have assessed cHT by using isogeneic cells in a rat model. However, to achieve clinical translation it is necessary to (1) understand how the cHT dose and timing of implantation affects safety and bone health, (2) identify suitable human, allogeneic, and/or xenogeneic cell sources for cHT and (3) understand how cHT achieves its effects at both a cellular and systemic level. We will also determine the maximum duration of time that cHT can secrete ovarian hormones and regulate other hormones of the HPO axis. The proposed experiments will be conducted primarily in a rat surgical model of ovarian failure to induce osteoporosis (ovariectomy; ovx). However, as a first step towards clinical translation we also propose the use of a larger, rabbit model of ovarian failure and osteoporosis (ovx with glucocorticoid delivery). cHT will be directly compared to pHT and to alendronate, a bisphosphonate drug widely prescribed to treat osteoporosis. We will also compare cHT (and controls) to untreated ovx animals and to rats that have not received an ovx (healthy, age-matched rats). We will propose three experimental aims: Aim 1. Determine effect of isogeneic cHT dose and implant timing on HPO hormone levels, safety and bone health in rat ovx model. Aim 2. Assess HPO hormone levels, immune response, safety and bone health in two ovarian failure models of osteoporosis with isogeneic, allogeneic or human cHT treatment. Aim 3. Determine differential gene expression (DGE) profiles for isogeneic, allogeneic, and human cHT to identify key functional pathways and any time-dependent changes in vitro and in a rat ovx model. Completion of these studies will demonstrate the safety and efficacy of these tissue-engineered constructs as a potential treatment for osteoporosis and provide information on their mechanisms of action.

持续调节下丘脑 - 垂体 - 卵巢激素与组织 - 工程卵巢构造作为女性骨质疏松症的治疗 项目摘要/摘要 雌性受骨质疏松症和骨质疏松性骨折的影响不成比例。在美国，大约 有4000万妇女有或有患骨质疏松症的风险，而200万人中的大多数（约60％） 女性发生骨质疏松骨折（每年直接费用> 200亿美元）。女性有1英寸2英寸的一生 骨质疏松性骨折的机会和骨质疏松症的发生率高4倍。钥匙的基础 原因是由于绝经或其他导致卵巢激素损失而导致的卵巢衰竭 下丘脑 - 卵巢 - 垂体（HPO）轴的伴随破坏。 直到2002年，传统的药物骑马疗法（PHT）被广泛使用，因为它的感知能力 降低骨质疏松症的风险，重要的是骨质疏松性骨折。妇女健康计划（WHI）已验证 这种长期以来的信念表明，PHT减少了骨质疏松骨折的入射。但是，也 表明骑马疗法的风险，例如心血管疾病和某些类型的癌症， 超过降低骨质疏松症水平的好处。后续研究表明风险可能更高 在60岁以上的女性和/或10岁以上的妇女中，这表明骑马酮 如果以较低的安全剂量和合适的递送平台给予治疗，则可能仍然有效。 我们提出仿生的卵巢细胞构建体作为雌激素递送的组织工程方法（细胞） 马龙治疗； CHT）治疗骨质疏松症。指导这项研究的假设是CHT可以 与PHT或PHT等药物相比，获得更好的骨骼健康和安全结果 双膦酸盐，在卵巢衰竭中，因为它模仿了天然卵巢结构和功能。 我们的CHT方法使用两种关键卵巢细胞类型（颗粒和theca细胞）的空间排列 模仿天然卵巢卵泡结构，同时避免了整个卵巢组织封装的陷阱，例如 卵母细胞（鸡蛋）驱动和随时间的功能丧失。我们的系统已经实现了持续和身体的实现 相关的卵巢骑马分泌水平。我们已经证明了封装的卵巢细胞 参与HPO轴，这可以通过刺激马酮（FSH）和黄体生成的调节证明 马龙（LH）从前垂体中水平。我们的CHT结构之间存在根本差异 和本地卵巢（例如，我们的结构中缺乏卵母细胞），但卵巢，垂体和可能的下丘脑 骑马的监管方式类似于伏雄前衰竭状态。这些基于细胞的构造也 释放其他由本地卵巢分泌的骑兵，但在PHT中不存在。更重要的是，CHT 以比PHT更安全的方式实现了有益的骨骼结局。 这项研究的长期目标是开发一种基于细胞的疗法，可以在人类中用于人类 预防与女性卵巢功能丧失相关的骨质疏松症。在我们的初步研究中，我们有 通过在大鼠模型中使用同系细胞评估CHT。但是，要实现临床翻译，有必要 （1）了解植入的CHT剂量和时间如何影响安全性和骨骼健康，（2）确定合适的 人类，同种异体和/或异种细胞来源，（3）了解CHT如何在 均和全身水平。我们还将确定CHT可以秘密的最长持续时间 卵巢骑兵并调节HPO轴的其他骑手。 提出的实验将主要在卵巢失效的大鼠手术模型中进行 骨质疏松症（卵巢切除术； OVX）。但是，作为临床翻译的第一步，我们还建议使用 卵巢衰竭和骨质疏松症的较大的兔模型（具有糖皮质激素递送的OVX）。 CHT将直接 与PHT和Alendronate相比，双膦酸盐药物被广泛处方用于治疗骨质疏松症。我们将 还将CHT（和对照组）与未经治疗的OVX动物以及未接受OVX（健康，健康的大鼠）进行比较（健康， 年龄匹配的大鼠）。我们将提出三个实验目标： 目标1。确定异质化CHT剂量和植入时间对HPO激素水平，安全性和 大鼠OVX模型中的骨骼健康。 目标2。评估两个卵巢衰竭的HPO赛马水平，免疫响应，安全性和骨骼健康 骨质疏松症的模型，以及同种异体，同种异体或人类CHT治疗的模型。 AIM 3。确定异种，同种异体和人类CHT的差异基因表达（DGE）谱 识别关键功能途径以及在体外和大鼠OVX模型中的任何时间依赖性变化。 这些研究的完成将证明这些组织工程结构的安全性和效率 骨质疏松症的潜在治疗方法，并提供有关其作用机理的信息。