Mechanisms of Tissue Destruction in Cholesteatoma

描述（由申请人提供）：胆脂瘤是一种上皮肿瘤样生长，起源于鼓膜，通常是慢性中耳炎的后遗症。这种病变的进行性通常会导致中耳和内耳结构通过骨吸收的破坏。这种破坏导致听力损失、前庭功能障碍和颅内并发症。目前对这种疾病的治疗仅限于手术根除，必须经常重复。慢性感染是中耳炎的另一个标志，因此我们一直对导致破骨细胞活化的炎症信号通路感兴趣。我们最近确定了一氧化氮合酶1（NOS 1或神经元NOS）在破骨细胞活化中的作用。我们现在希望通过确定NOS1被激活的机制和保持一氧化氮产生的内源性调节剂来进一步探索这一作用。为此，我们概述了以下具体目标。具体目标1：定位介导破骨细胞活性的NOS1的细胞来源，并定位NOS1的潜在内源性调节剂。具体目标二：确定NOS1的激活机制，并评估各种调节剂控制一氧化氮产生的效率，并最终在中耳炎模型中控制破骨细胞的骨吸收活性。神经元通过将NOS1功能性地连接到NMDA型谷氨酸受体来控制NOS1活化。有趣的是，破骨细胞也具有功能性谷氨酸受体，包括NMDA亚型，但其功能尚不清楚。此外，谷氨酸受体拮抗剂抑制破骨细胞密封骨的能力，这是再吸收活性的先决条件。来自具有NOS 1基因的靶向缺失的小鼠的破骨细胞表现出类似的表型。这一巧合提供了有趣的证据，表明这两个观察结果是相关的，破骨细胞可能使用与神经元相同的机制来调节NOS 1激活。这些研究提供了重要的洞察破骨细胞活性的控制，并可能导致药物干预的发展，将取代手术治疗这种衰弱性疾病的需要。

DESCRIPTION (provided by applicant): Cholesteatomas are epithelial tumor-like growths that arise from the tympanic membrane, typically as a sequela of chronic otitis media. The progressive nature of this lesion commonly results in the destruction of middle and inner ear structures via osteoclastic bone resorption. This destruction leads to hearing loss, vestibular dysfunction and intracranial complications. Current management of this disease is limited to surgical eradication, which must often be repeated. Chronic infection is another hallmark of the cholesteatoma and we have therefore been interested in the inflammatory signaling pathways that lead to osteoclast activation. We recently established a role for nitric oxide synthase 1 (NOS1 or neuronal NOS) in osteoclast activation. We now wish to explore this role further by identifying the mechanisms by which NOS1 is activated and the endogenous modulators that keep nitric oxide production in check. To this end, we have outlined the following specific aims. Specific Aim 1: To localize the cellular source(s) of NOSl that mediate osteoclast activity and to also localize potential endogenous modulators of NOSl. Specific Aim 2: To identify the mechanism of activation for NOSl and assess the efficiency by which various modulators can control nitric oxide production and ultimately the bone resorbing activity of osteoclasts in models of cholesteatoma. Neurons control NOSl activation by functionally linking it to the NMDA type of glutamate receptor. Interestingly, osteoclasts also have functional glutamate receptors, including the NMDA subtype, but their function is not well understood. Further, glutamate receptor antagonists inhibit the ability of the osteoclast to seal to the bone, a prerequisite to resorption activity. Osteoclasts from mice with targeted deletions of the NOS1 gene exhibit a similar phenotype. This coincidence offers intriguing evidence that these two observations are related and that osteoclasts may use the same mechanisms that neurons do to modulate NOS1 activation. These studies offer important insight into the control of osteoclast activity in cholesteatoma and may lead to the development of pharmacologic interventions that would replace the need for surgery in the treatment of this debilitating disease.