Clarisse Eveno1, Patricia Hainaud2, Aurore Rampanou2, Philippe Bonnin3, Sana Bakhouche2, Evelyne Dupuy2, Jean-Olivier Contreres2, Marc Pocard4. 1. Univ Paris Diderot, Sorbonne Paris Cité, INSERM U965, Angiogenèse et recherche translationnelle, Hôpital Lariboisière, Paris, France; Univ Paris Diderot, Sorbonne Paris Cité, AP-HP, Hôpital Lariboisière, Chirurgie Digestive, Paris, France. 2. Univ Paris Diderot, Sorbonne Paris Cité, INSERM U965, Angiogenèse et recherche translationnelle, Hôpital Lariboisière, Paris, France. 3. Univ Paris Diderot, Sorbonne Paris Cité, INSERM U965, Angiogenèse et recherche translationnelle, Hôpital Lariboisière, Paris, France; Univ Paris Diderot, Sorbonne Paris Cité, AP-HP, Hôpital Lariboisière, Physiologie Clinique Explorations Fonctionnelles, Paris, France. 4. Univ Paris Diderot, Sorbonne Paris Cité, INSERM U965, Angiogenèse et recherche translationnelle, Hôpital Lariboisière, Paris, France; Univ Paris Diderot, Sorbonne Paris Cité, AP-HP, Hôpital Lariboisière, Chirurgie Digestive, Paris, France. Electronic address: marc.pocard@inserm.fr.
Abstract
BACKGROUND: An interaction between tumor cells and the microenvironment, as well as the development of angiogenesis, are required to form liver metastases (LMs). MATERIAL AND METHODS: Immunofluorescence detection of α-smooth muscle actin, desmin, Ki67, laminin, and CD31 was used to analyze the kinetics of tumor angiogenesis determinants, especially the contribution of hepatic stellate cells (HSCs) to angiogenesis in hepatic metastasis produced by intrasplenically injected LS174 colorectal cancer cells. Immunostaining was performed at various times (days 9, 14, 28, and 39). RESULTS: At the earliest stage, micrometastases consisted of proliferating cancer cells, a well-organized network of activated HSCs and laminin deposits. No vascular network was observed. As the LMs grew in size, an organized vascular network appeared; the laminin network colocalized with CD31 immunostaining. At the later stages, all the immunostained markers became peripheral as a central necrosis developed. Purified activated HSCs isolated from transgenic mice livers developing hepatocellular carcinoma secreted laminin and showed enhanced human umbilical vein EC network formation in a Matrigel assay. In a coinjection LM experiment, activated HSCs enhanced the metastatic process. Moreover, colorectal LMs from six patients were analyzed, and a pattern of marker distribution similar to the coinjection experiment was found in human LMs. CONCLUSIONS: For the first time, our results show that HSCs play a crucial role in organizing and accelerating the progression of metastasis in modulating the prometastatic niche, interacting with colorectal cancer cell recruitment, and the organization of angiogenesis during colorectal LM development. Therefore, HSCs may be an early therapeutic target in colorectal cancer therapies.
BACKGROUND: An interaction between tumor cells and the microenvironment, as well as the development of angiogenesis, are required to form liver metastases (LMs). MATERIAL AND METHODS: Immunofluorescence detection of α-smooth muscle actin, desmin, Ki67, laminin, and CD31 was used to analyze the kinetics of tumor angiogenesis determinants, especially the contribution of hepatic stellate cells (HSCs) to angiogenesis in hepatic metastasis produced by intrasplenically injected LS174 colorectal cancer cells. Immunostaining was performed at various times (days 9, 14, 28, and 39). RESULTS: At the earliest stage, micrometastases consisted of proliferating cancer cells, a well-organized network of activated HSCs and laminin deposits. No vascular network was observed. As the LMs grew in size, an organized vascular network appeared; the laminin network colocalized with CD31 immunostaining. At the later stages, all the immunostained markers became peripheral as a central necrosis developed. Purified activated HSCs isolated from transgenic mice livers developing hepatocellular carcinoma secreted laminin and showed enhanced human umbilical vein EC network formation in a Matrigel assay. In a coinjection LM experiment, activated HSCs enhanced the metastatic process. Moreover, colorectal LMs from six patients were analyzed, and a pattern of marker distribution similar to the coinjection experiment was found in human LMs. CONCLUSIONS: For the first time, our results show that HSCs play a crucial role in organizing and accelerating the progression of metastasis in modulating the prometastatic niche, interacting with colorectal cancer cell recruitment, and the organization of angiogenesis during colorectal LM development. Therefore, HSCs may be an early therapeutic target in colorectal cancer therapies.
Authors: Amber L Simpson; Alexandre Doussot; John M Creasy; Lauryn B Adams; Peter J Allen; Ronald P DeMatteo; Mithat Gönen; Nancy E Kemeny; T Peter Kingham; Jinru Shia; William R Jarnagin; Richard K G Do; Michael I D'Angelica Journal: Ann Surg Oncol Date: 2017-05-30 Impact factor: 5.344