A genetic strategy for combined screening and localized imaging of breast cancer.
A genetic strategy for combined screening and localized imaging of breast cancer.
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DOI：
10.1007/s11307-010-0377-y
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发表时间：
2011-06
影响因子：
3.1
通讯作者：
Zinn, Kurt R.
中科院分区：
文献类型：
--
作者：
Warram, Jason M.;Borovjagin, Anton V.;Zinn, Kurt R.
需要改进乳腺癌的早期检测，因为目前的成像方法缺乏检测小肿瘤和评估其疾病表型的敏感性。为了解决这一问题，构建了双报告腺病毒载体(Ad5/3-Id1-SEAP-Id1-mCherry)，该载体由肿瘤特异性Id1启动子驱动基于血液的筛查报告(分泌型胚胎碱性磷酸酶，SEAP)和荧光成像报告(MCherry)的表达。该诊断系统评估了其在各种侵袭性表型的乳腺癌细胞株上的筛选潜力。用免疫印迹法检测报告基因的表达，并与启动子水平的表达相关联。用荧光素酶感染性分析对腺病毒受体的表达与报告表达进行正常化。将Ad5/3-Id1-SEAP-Id1-mCherry感染的MDA-MB-231细胞和未感染的细胞联合移植到裸鼠的乳房脂肪垫内，以重复低剂量的肿瘤输送。监测ID1驱动的SEAP表达和mCherry成像以验证诊断的敏感性和有效性。被感染的乳腺癌细胞株在感染后2天时，培养上清液中SEAP的水平是本底的10倍。将Ad5/3-Id1-SEAP-Id1-mCherry感染细胞(MOI=10)植入裸鼠体内，当肿瘤中只有2.5%的肿瘤含有感染细胞时，血清SEAP水平比基线增加了14倍。对于mCherry报告基因也发现了这种强烈的反应，在植入后第2天的肿瘤移植瘤中明显可见。这一诊断系统将筛查与成像相结合，用于乳腺癌的早期检测和监测，可以很容易地扩展到其他记者/模式和癌症靶向方法。在基因、癌症特异性机制中结合筛查和成像，可以对乳腺癌进行灵敏的多模式检测和定位。
Improvements are needed for the early detection of breast cancer, as current imaging methods lack sensitivity to detect small tumors and assess their disease phenotype. To address this issue the dual reporter adenoviral vector (Ad5/3-Id1-SEAP-Id1-mCherry) was produced with a cancer specific Id1 promoter driving expression of a blood-based screening reporter (secreted embryonic alkaline phosphatase, SEAP) and a fluorescent imaging reporter (mCherry). This diagnostic system was assessed for its screening potential on breast cancer cell lines of various aggressive phenotypes. Reporter expression was measured and correlated with promoter level expression using western blot. Adenovirus receptor expression was normalized against reporter expression with luciferase infectivity assays. Ad5/3-Id1-SEAP-Id1-mCherry infected MDA-MB-231 cells combined with uninfected cells were implanted into the mammary fat pad of athymic nude mice to recapitulate low dose tumor delivery. Id1 driven SEAP expression and mCherry imaging were monitored to validate diagnostic sensitivity and efficacy. Infected breast cancer cell lines displayed SEAP levels in the media that were 10-fold above background by 2 days after infection. Ad5/3-Id1-SEAP-Id1-mCherry infected cells (MOI=10) implanted in athymic nude mice demonstrated a 14-fold increase in serum SEAP levels over baseline when as little as 2.5% of the tumor contained infected cells. This robust response was also found for the mCherry reporter which was clearly visible in tumor xenografts on day 2 post implantation. This diagnostic system that combines screening with imaging for early detection and monitoring of breast cancer can be easily extended to other reporters/modalities and cancer-targeting methods. Combining screening with imaging in a genetic, cancer-specific mechanism allows sensitive multi-modal detection and localization of breast cancer.
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影响因子：
120.7
作者：
Kerlikowske, K;Grady, D;Ernster, V
通讯作者：
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