Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, United States
Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, United States.
Dis Model Mech. 2018 Jul 30;11(7):dmm034793. doi: 10.1242/dmm.034793.
Syngeneic, immunocompetent allograft tumor models recapitulate important aspects of the tumor microenvironment and have short tumor latency with predictable growth kinetics, making them useful for trialing novel therapeutics. Here, we describe surgical techniques for orthotopic and heterotopic pancreatic ductal adenocarcinoma (PDAC) tumor implantation and characterize phenotypes based on implantation site.Mice (=8 per group) were implanted with 10 cells in the pancreas or flank. Hy15549 and Han4.13 cell lines were derived from primary murine PDAC (Ptf1-Cre; LSL-KRAS-G12D; Trp53 Lox/+) on C57BL/6 and FVB strains, respectively. Single-cell suspension and solid tumor implants were compared. Tumors were treated with two intravenous doses of FOLFIRINOX and responses evaluated.All mice developed pancreatic tumors within 7 days. Orthotopic tumors grew faster and larger than heterotopic tumors. By 3 weeks, orthotopic mice began losing weight, and showed declines in body condition requiring euthanasia starting at 4 weeks. Single-cell injection into the pancreas had near 100% engraftment, but solid tumor implant engraftment was ∼50% and was associated with growth restriction. Orthotopic tumors were significantly more responsive to intravenous FOLFIRINOX compared with heterotopic tumors, with greater reductions in size and increased apoptosis. Heterotopic tumors were more desmoplastic and hypovascular. However, drug uptake into tumor tissue was equivalent regardless of tumor location or degree of fibrosis, indicating that microenvironment differences between heterotopic and orthotopic tumors influenced response to therapy.Our results show that orthotopic and heterotopic allograft locations confer unique microenvironments that influence growth kinetics, desmoplastic response and angiogenesis. Tumor location influences chemosensitivity to FOLFIRINOX and should inform future preclinical trials.This article has an associated First Person interview with the first author of the paper.
同种异体、免疫活性的移植瘤模型再现了肿瘤微环境的重要方面，并且潜伏期短，生长动力学可预测，因此可用于试验新的治疗方法。在这里，我们描述了胰腺和异位胰腺导管腺癌（PDAC）肿瘤原位和异位种植的手术技术，并根据种植部位对表型进行了特征描述。每组（n=8）小鼠分别在胰腺或侧腹部植入 10 个细胞。Hy15549 和 Han4.13 细胞系分别来源于 C57BL/6 和 FVB 品系上带有 Ptf1-Cre；LSL-KRAS-G12D；Trp53 Lox/+ 的原发性小鼠 PDAC（Ptf1-Cre；LSL-KRAS-G12D；Trp53 Lox/+）。比较了单细胞悬液和实体瘤植入物。用两剂静脉注射 FOLFIRINOX 进行治疗，并评估了反应。所有小鼠在 7 天内均发展出胰腺肿瘤。原位肿瘤的生长速度和体积均大于异位肿瘤。到第 3 周，原位小鼠开始减重，并且从第 4 周开始身体状况下降，需要安乐死。将细胞悬液直接注射到胰腺中，几乎 100%的细胞都能植入，但实体瘤植入的细胞仅为 50%，并且与生长受限有关。与异位肿瘤相比，原位肿瘤对静脉注射 FOLFIRINOX 更为敏感，肿瘤体积缩小和凋亡增加更为明显。异位肿瘤的纤维化程度更高，血管生成较少。但是，无论肿瘤位置或纤维化程度如何，肿瘤组织中的药物摄取量均相等，这表明异位和原位肿瘤之间的微环境差异影响了对治疗的反应。我们的结果表明，同种异体的原位和异位种植部位赋予了独特的微环境，影响了生长动力学、纤维母细胞反应和血管生成。肿瘤位置影响 FOLFIRINOX 的化疗敏感性，这应该为未来的临床前试验提供信息。本文附有该论文第一作者的第一人称采访。
