胰腺癌中KRASG12D的选择性上皮表达驱动导管增殖和结缔组织增生,并伴有免疫反应。

IF 3.9 2区 综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES
Carlos P Jara, Al-Murtadha Al-Gahmi, Audrey Lazenby, Michael A Hollingsworth, Mark A Carlson
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引用次数: 0

摘要

胰腺导管腺癌(Pancreatic ductal adencarcinoma, PDAC)仍然是肿瘤学领域的一个巨大挑战,其特点是死亡率高,主要归因于诊断延迟和肿瘤微环境的复杂性。模拟胰腺上皮转化的创新为PDAC的发病机制和潜在的治疗策略提供了有价值的见解。我们采用猪(Oncopig)模型,利用Ad-K8-Cre腺病毒载体,研究了不同剂量(107至1010 pfu)对胰腺上皮细胞的影响。该载体的表达由角蛋白8启动子驱动,将cre -重组酶特异性地传递到上皮细胞。导管内胰腺注射转基因Oncopigs (LSL-KRASG12D-TP53R167H),注射后2个月进行组织学评估。通过角蛋白-8表达和cre -重组酶活性验证腺病毒载体的特异性。我们通过角蛋白8和CAM5.2染色证实,Ad-K8-Cre腺病毒载体主要靶向大、小胰管内壁的导管上皮细胞。较高剂量导致显著的组织形态改变,包括萎缩和淋巴结肿大。显微镜检查显示浓度依赖性导管上皮增生,细胞异型,化生和间质改变。免疫组织化学证实转基因表达。通过vimentin、α-SMA和Masson三色染色,可见明显的胶原形成反应,表明胶原沉积在进行性,特别是在较高的载体剂量下。我们的研究表明,在Oncopig模型中,Ad-K8-Cre在诱导胰腺上皮增生和可能的肿瘤形成方面存在明显的剂量-反应关系。所有剂量的载体均诱导上皮细胞增殖;高剂量也会引起间质改变、化生和可能的肿瘤转化。这些发现强调了在上皮肿瘤的大型动物模型中致癌基因的位点特异性激活的潜力,具有诱导与人类PDAC相似的基质改变的能力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Selective epithelial expression of KRASG12D in the Oncopig pancreas drives ductal proliferation and desmoplasia that is accompanied by an immune response.

Pancreatic ductal adenocarcinoma (PDAC) remains a formidable challenge in oncology, characterized by a high mortality rate, largely attributable to delayed diagnosis and the intricacies of its tumor microenvironment. Innovations in modeling pancreatic epithelial transformation provide valuable insights into the pathogenesis and potential therapeutic strategies for PDAC. We employed a porcine (Oncopig) model, utilizing the Ad-K8-Cre adenoviral vector, to investigate the effects of variable doses (107 to 1010 pfu) on pancreatic epithelial cells. This vector, the expression from which being driven by a Keratin-8 promoter, will deliver Cre-recombinase specifically to epithelial cells. Intraductal pancreatic injections in transgenic Oncopigs (LSL-KRASG12D-TP53R167H) were performed with histologically based evaluation at 2 months post-injection. Specificity of the adenoviral vector was validated through Keratin-8 expression and Cre-recombinase activity. We confirmed that the Ad-K8-Cre adenoviral vector predominantly targets ductal epithelial cells lining both large and small pancreatic ducts, as evidenced by Keratin 8 and CAM5.2 staining. Higher doses resulted in significant tissue morphology changes, including atrophy, and enlarged lymph nodes. Microscopic examination revealed concentration-dependent proliferation of the ductal epithelium, cellular atypia, metaplasia, and stromal alterations. Transgene expression was confirmed with immunohistochemistry. Desmoplastic responses were evident through vimentin, α-SMA, and Masson's trichrome staining, indicating progressive collagen deposition, particularly at the higher vector doses. Our study suggests a distinct dose-response relationship of Ad-K8-Cre in inducing pancreatic epithelial proliferation and possible neoplasia in an Oncopig model. All doses of the vector induced epithelial proliferation; the higher doses also produced stromal alterations, metaplasia, and possible neoplastic transformation. These findings highlight the potential for site-specific activation of oncogenes in large animal models of epithelial tumors, with the ability to induce stromal alterations reminiscent of human PDAC.

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来源期刊
Scientific Reports
Scientific Reports Natural Science Disciplines-
CiteScore
7.50
自引率
4.30%
发文量
19567
审稿时长
3.9 months
期刊介绍: We publish original research from all areas of the natural sciences, psychology, medicine and engineering. You can learn more about what we publish by browsing our specific scientific subject areas below or explore Scientific Reports by browsing all articles and collections. Scientific Reports has a 2-year impact factor: 4.380 (2021), and is the 6th most-cited journal in the world, with more than 540,000 citations in 2020 (Clarivate Analytics, 2021). •Engineering Engineering covers all aspects of engineering, technology, and applied science. It plays a crucial role in the development of technologies to address some of the world''s biggest challenges, helping to save lives and improve the way we live. •Physical sciences Physical sciences are those academic disciplines that aim to uncover the underlying laws of nature — often written in the language of mathematics. It is a collective term for areas of study including astronomy, chemistry, materials science and physics. •Earth and environmental sciences Earth and environmental sciences cover all aspects of Earth and planetary science and broadly encompass solid Earth processes, surface and atmospheric dynamics, Earth system history, climate and climate change, marine and freshwater systems, and ecology. It also considers the interactions between humans and these systems. •Biological sciences Biological sciences encompass all the divisions of natural sciences examining various aspects of vital processes. The concept includes anatomy, physiology, cell biology, biochemistry and biophysics, and covers all organisms from microorganisms, animals to plants. •Health sciences The health sciences study health, disease and healthcare. This field of study aims to develop knowledge, interventions and technology for use in healthcare to improve the treatment of patients.
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