TY - JOUR
T1 - Microarchitectural mimicking of stroma-induced vasculature compression in pancreatic tumors using a 3D engineered model
AU - Heinrich, Marcel Alexander
AU - Uboldi, Irene
AU - Kuninty, Praneeth Reddy
AU - Ankone, Marc J.K.
AU - van Baarlen, Joop
AU - Zhang, Yu Shrike
AU - Jain, Kartik
AU - Prakash, Jai
N1 - Funding Information:
To further confirm the high viability of cells we measured the release of lactate dehydrogenase (LDH) into the culture as indicator of cytotoxicity. Here, we first investigated the release of LDH after incubation for 24 h in fresh culture medium (Fig. 2G and Fig. S3, Supporting Information). We found that while PSCs in collagen show a slightly higher release of LDH, indicating higher toxicity, the overall measured values are very low being roughly 10x lower compared to the positive control displaying the maximum release of LDH possible. This indicates an overall high viability of PSCs in both, collagen and Col/Fib, despites small differences. Additionally we investigated the cumulative release of LDH for a total duration of 7 days while only refreshing parts of the medium (Fig. 2H and Fig. S3, Supporting Information). We found that after 7 days the concentration of LDH in the medium doubled, however, still remained significantly low compared to the positive control again indicating an overall high viability of PSCs for the culture duration. Interestingly, also here we observed a slightly higher release of LDH for collagen gels at day 7 post-preparation which for both conditions might be based on the contraction of the hydrogels entrapping parts of the LDH.The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Jai Prakash reports financial support was provided by Dutch Research Council. Kartik Jain reports financial support was provided by Dutch Research Council. Jai Prakash reports a relationship with ScarTec Therapeutics that includes: equity or stocks. Jai Prakash has patent #AU2016340763B2 issued to University of Twente/ScarTec Therapeutics. Jai Prakash has patent #EP3365353A1 pending to University of Twente/ScarTec Therapeutics. Other authors declare no conflict of interest.
Funding Information:
Marcel Alexander Heinrich: contributed equally to this work as primary author, conceived the study, designed experiments, and co-wrote the manuscript, and, performed the in vitro experiments. Irene Uboldi: contributed equally to this work as primary author, performed the in vitro experiments. Praneeth Reddy Kuninty: performed the animal studies and the immunohistochemical staining of tissue sections. Marc J.K. Ankone: performed the mechanical analysis of the hydrogels. Joop van Baarlen: provided the sections from human PDAC patients. Yu Shrike Zhang: helped in discussions on the project and reviewed the manuscript. Kartik Jain: performed the CFD simulations and supervised the fluid mechanical aspects. Jai Prakash: conceived the study, designed experiments, and co-wrote the manuscript. All authors reviewed the manuscript. Correspondence and requests for materials should be addressed to Jai Prakash. The authors acknowledge funding from the Dutch Technology Foundation and the Dutch Cancer Society (STW/ KWF; project no. 15204 to J.P.). Compute resources on the Dutch national supercomputer Cartesius were provided by SURFSara through NWO grant 2019/ENW/00768083.
Funding Information:
The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Jai Prakash reports financial support was provided by Dutch Research Council . Kartik Jain reports financial support was provided by Dutch Research Council. Jai Prakash reports a relationship with ScarTec Therapeutics that includes: equity or stocks. Jai Prakash has patent #AU2016340763B2 issued to University of Twente/ScarTec Therapeutics. Jai Prakash has patent #EP3365353A1 pending to University of Twente/ScarTec Therapeutics. Other authors declare no conflict of interest.
Publisher Copyright:
© 2022 The Authors
Financial transaction number:
2500075460
PY - 2023/4
Y1 - 2023/4
N2 - Fibrotic tumors, such as pancreatic ductal adenocarcinoma (PDAC), are characterized for high desmoplastic reaction, which results in high intra-tumoral solid stress leading to the compression of blood vessels. These microarchitectural alterations cause loss of blood flow and poor intra-tumoral delivery of therapeutics. Currently, there is a lack of relevant in vitro models capable of replicating these mechanical characteristics and to test anti-desmoplastic compounds. Here, a multi-layered vascularized 3D PDAC model consisting of primary human pancreatic stellate cells (PSCs) embedded in collagen/fibrinogen (Col/Fib), mimicking tumor tissue within adjunct healthy tissue, is presented to study the fibrosis-induced compression of vasculature in PDAC. It is demonstrated how the mechanical and biological stimulation induce PSC activation, extracellular matrix production and eventually vessel compression. The clinical relevance is confirmed by correlating with patient transcriptomic data. Furthermore, the effects of gradual vessel compression on the fluid dynamics occurring within the channel is evaluated in silico. Finally, it is demonstrated how cancer-associated fibroblast (CAF)-modulatory therapeutics can inhibit the cell-mediated compression of blood vessels in PDAC in vitro, in silico and in vivo. It is envisioned that this 3D model is used to improve the understanding of mechanical characteristics in tumors and for evaluating novel anti-desmoplastic therapeutics.
AB - Fibrotic tumors, such as pancreatic ductal adenocarcinoma (PDAC), are characterized for high desmoplastic reaction, which results in high intra-tumoral solid stress leading to the compression of blood vessels. These microarchitectural alterations cause loss of blood flow and poor intra-tumoral delivery of therapeutics. Currently, there is a lack of relevant in vitro models capable of replicating these mechanical characteristics and to test anti-desmoplastic compounds. Here, a multi-layered vascularized 3D PDAC model consisting of primary human pancreatic stellate cells (PSCs) embedded in collagen/fibrinogen (Col/Fib), mimicking tumor tissue within adjunct healthy tissue, is presented to study the fibrosis-induced compression of vasculature in PDAC. It is demonstrated how the mechanical and biological stimulation induce PSC activation, extracellular matrix production and eventually vessel compression. The clinical relevance is confirmed by correlating with patient transcriptomic data. Furthermore, the effects of gradual vessel compression on the fluid dynamics occurring within the channel is evaluated in silico. Finally, it is demonstrated how cancer-associated fibroblast (CAF)-modulatory therapeutics can inhibit the cell-mediated compression of blood vessels in PDAC in vitro, in silico and in vivo. It is envisioned that this 3D model is used to improve the understanding of mechanical characteristics in tumors and for evaluating novel anti-desmoplastic therapeutics.
KW - 3D in vitro model
KW - Cancer-associated fibroblasts
KW - Collagen
KW - Computational flow dynamics
KW - Fibrinogen
KW - Pancreatic ductal adenocarcinoma
UR - http://www.scopus.com/inward/record.url?scp=85138668023&partnerID=8YFLogxK
U2 - 10.1016/j.bioactmat.2022.09.015
DO - 10.1016/j.bioactmat.2022.09.015
M3 - Article
SN - 2452-199X
VL - 22
SP - 18
EP - 33
JO - Bioactive Materials
JF - Bioactive Materials
ER -
