(PQB5) Does the timing of Pten and Rb1 mutation affect prostate cancer phenotypes

(PQB5) Pten 和 Rb1 突变的时间是否影响前列腺癌表型

• 批准号: 8587206
• 负责人: DAVID W. GOODRICH
• 金额: $ 22.16万
• 依托单位: ROSWELL PARK CANCER INSTITUTE CORP
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8587206
• 关键词: Address; Affect; Alleles; Androgens; Behavior; Binding; Breeding; Cancer Model; Cancer Patient; Cancer Prognosis; Cells; Chimeric Proteins; Data; Disease; Ensure; Epigenetic Process; Estrogen Receptors; Future; Gene Mutation; Genes; Genetic; Genetic Engineering; Genetic Predisposition to Disease; Genetic Recombination; Genetic Transcription; Genetically Engineered Mouse; Goals; Histologic; Human; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of prostate; Mediating; Methodology; Mus; Mutation; Nature; PTEN gene; Pattern; Performance; Phenotype; Process; RB1 gene; Recurrence; Retinoblastoma; Site; Staging; System; Tamoxifen; Testing; Time; Tissues; Transgenes; Transgenic Mice; Translations; Variant; base; cancer cell; cancer initiation; cancer therapy; deprivation; design; expectation; human disease; in vivo; interest; lung small cell carcinoma; mouse model; novel; prostate cancer model; public health relevance; recombinase; response; tumor progression

DESCRIPTION (provided by applicant): Cancer progression is driven by an evolutionary process of genetic mutation, phenotypic variation, and selection. The genetic background of cancer cells is thus continuously changing. Genetic background will dictate the nature of future mutations that can be tolerated and selected for. The effects of specific new mutations will depend on the genetic background in which they occur. Thus the temporal order of mutations is expected to influence cancer phenotype. How the timing of mutation affects cancer phenotypes remains a largely unanswered question, however. Answering this provocative question (PQB5) is significant because cancer prognosis and effective personalized cancer treatment will depend not only on the nature of mutations present within a particular cancer, but also on the order in which they are acquired. Genetically engineered mouse cancer models have been vital in elucidating the genetic etiology of cancer, but have rarely been used to address PQB5. This is due to limitations inherent in commonly used genetic engineering methodology; when multiple mutations are created, the order of mutations is either unknown, is not controllable, or is not restricted to incipient cancer cells. A current barrier to progress is the availability of mouse cancer models that allow precise control over the timing and order of multiple genetic mutations. We propose to overcome this barrier by developing a novel mouse cancer model based on a lox-neo/stop-lox FlpO-ERT2 transgene. This transgene encodes an FlpO recombinase, ERT2 estrogen receptor fusion protein whose activity is tamoxifen inducible. The transgene is combined with tissue specific Cre transgenes, floxed alleles of the cancer initiating mutation, and frted alleles of the secondary mutation. Cre expression creates the cancer initiating mutation and removes the neo/stop cassette from lox-neo/stop-lox FlpO-ERT2, restricting its expression to initiated cancer cells. FlpO-ERT2 activity can then be induced at experimentally controlled times by tamoxifen administration, creating a secondary mutation by deleting frted gene alleles. This mouse model can be used to determine how 2 mutations occurring in a defined and controllable temporal order affect cancer phenotypes in vivo. We will use this mouse model to test whether the timing of Pten and Rb1 mutation alters prostate cancer phenotype. PTEN and RB1 mutation are common in human prostate cancer, with PTEN loss occurring early and RB1 loss occurring late. The late loss of RB1 is puzzling as its early loss is known to initiate other human cancers. It is unknown whether PTEN and RB1 loss cooperate to drive prostate cancer progression or whether the temporal pattern of mutation influences prostate cancer phenotype. We postulate that the timing of Pten and Rb1 mutation will alter prostate cancer phenotypes in vivo because the effects of these mutations are dependent on genetic background. Two specific aims are proposed to create the lox-neo/stop-lox FlpO-ERT2 allele, to use it to alter the timing of Pten and Rb1 mutation in the mouse prostate cancer model, and to characterize the effects of these mutations on prostate cancer phenotypes in vivo.

描述（由申请人提供）：癌症进展是由基因突变、表型变异和选择的进化过程驱动的。 因此，癌细胞的遗传背景不断变化。 遗传背景将决定未来可以耐受和选择的突变的性质。 特定新突变的影响将取决于它们发生的遗传背景。 因此，突变的时间顺序预计会影响癌症表型。 然而，突变的时间如何影响癌症表型仍然是一个很大程度上悬而未决的问题。 回答这个具有挑战性的问题（PQB5）具有重要意义，因为癌症预后和有效的个性化癌症治疗不仅取决于特定癌症中存在的突变的性质，还取决于它们获得的顺序。 基因工程小鼠癌症模型对于阐明癌症的遗传病因至关重要，但很少用于解决 PQB5。 这是由于常用基因工程方法固有的局限性；当产生多个突变时，突变的顺序要么是未知的，要么是不可控制的，要么不限于初期癌细胞。 目前进展的一个障碍是小鼠癌症模型的可用性，该模型可以精确控制多个基因突变的时间和顺序。 我们建议通过开发一种基于 lox-neo/stop-lox FlpO-ERT2 转基因的新型小鼠癌症模型来克服这一障碍。 该转基因编码 FlpO 重组酶、ERT2 雌激素受体融合蛋白，其活性是他莫昔芬诱导的。 该转基因与组织特异性Cre转基因、癌症起始突变的floxed等位基因和二次突变的frted等位基因组合。 Cre 表达产生癌症起始突变，并从 lox-neo/stop-lox FlpO-ERT2 中去除 neo/stop 盒，将其表达限制在起始癌细胞中。 然后可以通过给予他莫昔芬在实验控制的时间诱导FlpO-ERT2活性，通过删除frted基因等位基因产生二次突变。 该小鼠模型可用于确定以定义且可控的时间顺序发生的 2 个突变如何影响体内癌症表型。 我们将使用该小鼠模型来测试 Pten 和 Rb1 突变的时间是否会改变前列腺癌表型。 PTEN和RB1突变在人类前列腺癌中很常见，PTEN缺失发生在早期，RB1缺失发生在晚期。 RB1 的后期损失令人费解，因为它的早期损失是 已知会引发其他人类癌症。 目前尚不清楚 PTEN 和 RB1 缺失是否协同驱动前列腺癌进展，或者突变的时间模式是否影响前列腺癌表型。 我们假设 Pten 和 Rb1 突变的时间将改变体内前列腺癌表型，因为这些突变的影响取决于遗传背景。 提出了两个具体目标，即创建 lox-neo/stop-lox FlpO-ERT2 等位基因，用它来改变小鼠前列腺癌模型中 Pten 和 Rb1 突变的时间，并表征这些突变对体内前列腺癌表型的影响。