关键词：脑膜炎;生物荧光;乳腺癌;克隆;固化剂;转移;肿瘤;肿瘤;治疗;病毒
摘 要：Neoplastic meningitis is a severe complication of breast cancer for which there is no curative treatment at present. The project aims to develop a novel, safe and efficient therapy for neoplastic meningitis HSV-1 oncolysis therapy. For the first year of the award period we have propagated the HSV-1 replication conditional mutant (hrR3) with burst assays and generated three different virus preparations to establish viral titers of 1x10(7), 1x10(8) and 1x10(9) pfu/ml. The virus titers were determined by plaque assays performed on vero mono-layers. These virus titers will be used to study fractionated viruses particles. In addition we have commenced on additional work that will help study the disease progression by molecular imaging. In anticipation of establishing a mouse model of meningeal metastasis we have developed stable breast cancer cell lines that express bioluminescence and fluorescence markers for in vivo molecular imaging. Three cell lines (human breast cancer cell lines SkBr3 and MDA-231, and the mouse breast cancer 4T1) were transfected to express renilla luciferase (RLuc) and mCherry, both driven by the CMV promoter. Clones that were resistant to puromycin were selected to establish stable cell lines. These stable cell lines have been tested for (i) the replication efficiency HSV- 1, and (ii) their sensitivity to the HSV-1 oncolysis with in vitro bioluminescence. Based on these preliminary data we anticipate proposing the use of one of the cell lines to establish a mouse model of meningeal metastases. We will in due course approach DOD for approval of the animal protocol.