The Role of Protein Kinase D in Epidermal Tumorigenesis

蛋白激酶 D 在表皮肿瘤发生中的作用

• 批准号: 8241636
• 负责人: Wendy B Bollag
• 金额: --
• 依托单位: CHARLIE NORWOOD VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-10-01 至 2015-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8241636
• 关键词: Adverse effects; Age; Apoptosis; Artificial skin; Basal cell carcinoma; Biopsy; Cell Death; Cell Survival; Cells; Cessation of life; Chronic; Cicatrix; Cutaneous; DNA Damage; Data; Development; Diagnosis; Enzymes; Epidermis; Etiology; Excision; Gene Delivery; Gene Mutation; Harvest; Human; Human Volunteers; In Situ; In Vitro; Life; Light; Link; Location; Malignant Neoplasms; Malignant neoplasm of pancreas; Malignant neoplasm of prostate; Measures; Mitochondria; Monitor; Mus; Neoplasms; Operative Surgical Procedures; Pathway interactions; Patients; Photosensitivity; Population; Proliferating; Recurrence; Risk; Risk Factors; Role; Signaling Molecule; Skin; Skin Cancer; Skin Carcinoma; Skin Neoplasms; Squamous cell carcinoma; Stratum Basale; Sun Exposure; Testing; The Sun; Time; Topical application; Transgenic Mice; UV induced; UVB induced; Ultraviolet B Radiation; Ultraviolet Rays; United States; Veterans; adenoviral-mediated; cancer type; cell injury; cell type; chemotherapy; in vivo; inhibitor/antagonist; irradiation; keratinocyte; mutant; neoplastic; novel; novel therapeutics; overexpression; prevent; protein kinase D; reconstitution; repaired; research study; response; small molecule; therapy development; tumor; tumorigenesis; ultraviolet; ultraviolet damage; ultraviolet irradiation

DESCRIPTION (provided by applicant): The non-melanoma skin cancers (NMSCs), basal and squamous cell carcinoma, are the most common cancers, occurring more often than all cancers combined, with approximately 1 million new cases diagnosed per year. Although the NMSCs, in particular basal cell carcinomas (BCCs) are, for the most part, essentially curable, surgical treatment often results in scarring and disfigurement, and the risk of recurrence is high (the chance of a second occurrence is 35-50%). The major risk factors for the development of NMSCs are chronic sun exposure (ultraviolet irradiation) and age: thus, these cancers are a major problem in the veteran population. BCCs, which make up approximately 80% of the NMSCs, arise from basal keratinocytes in the epidermis of the skin. Our previous studies have demonstrated that protein kinase D (PKD) is up-regulated in keratinocytes in human BCCs. In addition, we recently demonstrated that PKD1 is activated upon exposure of mouse keratinocytes to ultraviolet B irradiation and protects these cells from UVB-induced apoptosis in vitro. These results suggest that UVB might select for cells with higher levels of pro-proliferative PKD1. Alternatively, active PKD1 may allow survival of UV-damaged cells. This ability of PKD1 to promote survival would be beneficial in preventing excessive apoptosis with low levels of UVB exposure, causing minimal DNA damage that can be repaired. However, if PKD1 allows survival of cells that have suffered irreparable UV-induced DNA damage, these keratinocytes with DNA mutations could continue to proliferate and form skin tumors. Thus, either a pro-proliferative or pro-survival mechanism could provide a means by which PKD1 could contribute to epidermal tumorigenesis. Moreover, these results suggest that small-molecule PKD inhibitors might be a viable therapy for the treatment of non-melanoma skin cancers. Because these inhibitors can be applied topically, with minimal systemic exposure, they could potentially be used with few side effects even if PKD1 has important roles in other cell types. In addition, PKD inhibitors have been proposed as possible novel therapies for treatment of pancreatic cancer. Therefore, an understanding of the role of PKD1 in the epidermis may be important in determining the possibility of epidermal side effects of systemic treatment with these agents. For example, inhibition of PKD1 in the skin could result in increased sun sensitivity, with ultraviolet light triggering massive apoptosis of the keratinocytes comprising the epidermis, in patients on PKD inhibitors for the treatment of internal cancers. In the experiments proposed here, we intend to determine the mechanisms by which ultraviolet B light- induced PKD1 activation protects keratinocytes from cell death, thereby promoting their survival. In addition we will extend our results obtained in mouse keratinocytes in vitro, to the mouse in vivo. Further, we will investigate the effect of ultraviolet B light on PKD1 activation in human keratinocytes in vitro, in reconstituted human skin in situ and in human volunteers in vivo. In particular, our aims are to: (1) test the hypothesis that UVB-activated PKD1 localizes to the mitochondria, at which location the enzyme protects keratinocytes from UV-induced apoptosis, (2) test the hypothesis that PKD1 is activated in human keratinocytes in vitro and reconstituted human epidermis in situ and promotes survival upon UVB exposure, (3) test the hypothesis that PKD1 is activated by UVB in mouse and human skin in vivo and (4) test the hypothesis that UVB selects for keratinocytes with higher PKD1 levels in vitro and in vivo. The results of these studies should allow the identification of novel signaling molecules, such as PKD1 or its downstream pathways, which may be targeted to treat and/or prevent NMSCs in veterans and the U.S. population as a whole. In addition, PKD inhibitors have been proposed as potential novel therapies for the treatment of other types of cancers, such as pancreatic cancer. Evidence concerning the role of PKD in the keratinocyte response to UVB may be important for predicting possible sun sensitivity in patients undergoing systemic chemotherapy with PKD inhibitors.

描述（由申请人提供）： 非黑色素瘤皮肤癌 (NMSC)、基底细胞癌和鳞状细胞癌是最常见的癌症，其发生频率比所有癌症的总和还要高，每年诊断出约 100 万新病例。虽然 NMSC，特别是基底细胞癌 (BCC) 在大多数情况下基本上是可以治愈的，但手术治疗往往会导致疤痕和毁容，并且复发风险很高（第二次发生的几率为 35-50%） ）。 NMSC 发展的主要危险因素是长期阳光照射（紫外线照射）和年龄：因此，这些癌症是退伍军人群体中的一个主要问题。 BCC 约占 NMSC 的 80%，源自皮肤表皮的基底角质形成细胞。我们之前的研究表明，人基底细胞癌的角质形成细胞中蛋白激酶 D (PKD) 上调。此外，我们最近证明，当小鼠角质形成细胞暴露于紫外线 B 照射时，PKD1 会被激活，并在体外保护这些细胞免受 UVB 诱导的细胞凋亡。这些结果表明 UVB 可能会选择具有较高水平的促增殖 PKD1 的细胞。或者，活性 PKD1 可以使受紫外线损伤的细胞存活。 PKD1 促进存活的能力将有利于防止低水平 UVB 暴露下的过度细胞凋亡，从而将可修复的 DNA 损伤降至最低。然而，如果 PKD1 允许遭受不可修复的紫外线诱导的 DNA 损伤的细胞存活，这些带有 DNA 突变的角质形成细胞可能会继续增殖并形成皮肤肿瘤。因此，促增殖或促存活机制都可以提供一种 PKD1 促进表皮肿瘤发生的方法。此外，这些结果表明小分子 PKD 抑制剂可能是治疗非黑色素瘤皮肤癌的可行疗法。由于这些抑制剂可以局部应用，全身暴露极少，因此即使 PKD1 在其他细胞类型中发挥重要作用，它们的副作用也很少。此外，PKD 抑制剂已被提议作为治疗胰腺癌的可能新疗法。因此，了解 PKD1 在表皮中的作用对于确定使用这些药物进行全身治疗可能产生表皮副作用可能很重要。例如，在使用 PKD 抑制剂治疗内科癌症的患者中，抑制皮肤中的 PKD1 可能会导致阳光敏感性增加，而紫外线会引发构成表皮的角质形成细胞的大量凋亡。 在此提出的实验中，我们打算确定紫外线 B 光诱导的 PKD1 激活保护角质形成细胞免遭细胞死亡的机制，从而促进其存活。此外，我们还将把在体外小鼠角质形成细胞中获得的结果扩展到小鼠体内。此外，我们将研究紫外线 B 光对体外人角质形成细胞、原位重建人皮肤以及人体志愿者体内 PKD1 激活的影响。具体而言，我们的目标是：(1) 检验 UVB 激活的 PKD1 定位于线粒体的假设，在该位置酶可保护角质形成细胞免受紫外线诱导的细胞凋亡，(2) 检验 PKD1 在人角质形成细胞中被激活的假设体外和原位重建人表皮并促进 UVB 暴露后的存活，(3) 测试 PKD1 在小鼠和人皮肤体内被 UVB 激活的假设，以及 (4)检验 UVB 选择体外和体内 PKD1 水平较高的角质形成细胞的假设。这些研究的结果应该能够识别新的信号分子，例如 PKD1 或其下游途径，这些分子可能有针对性地治疗和/或预防退伍军人和整个美国人口中的 NMSC。此外，PKD 抑制剂已被提议作为治疗其他类型癌症（例如胰腺癌）的潜在新疗法。关于 PKD 在角质形成细胞对 UVB 反应中的作用的证据可能对于预测接受 PKD 抑制剂全身化疗的患者可能的阳光敏感性很重要。