Engineered extracellular matrices reveal stiffness-mediated chemoresistance in patient-derived pancreatic cancer organoids

LeSavage, Bauer L.; Gilchrist, Aidan E.; Krajina, Brad A.; Karlsson, Kasper; Smith, Amber Rae; Karagyozova, Kremena; Klett, Katarina C.; Curtis, Christina; Kuo, Calvin J.; Heilshorn, Sarah C.

doi:10.1101/2022.04.22.488943

Cited by 10 publications

(24 citation statements)

References 72 publications

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“…This technology allows us to stiffen and soften a matrix and observe its effects on the same cell population longitudinally without the need for gel digestion and cell dissociation. 88 Here, using our platform technology, we found that, as expected, magneto-stiffening of the matrix promoted EMT and the hypoxic environment in tumor spheroids while it reduced drug sensitivity and tumor-killing effects. 12,89 However, during matrix softening on the same spheroid population, EMT was halted, and we observed a switch to MET based on the expression levels of E-/N-cadherin.…”

Section: Discussionsupporting

confidence: 72%

Dynamic Magneto-Softening of 3D Hydrogel Reverses Malignant Transformation of Cancer Cells and Enhances Drug Efficacy

Shou

Teo

et al. 2023

ACS Nano

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High extracellular matrix stiffness is a prominent feature of malignant tumors associated with poor clinical prognosis. To elucidate mechanistic connections between increased matrix stiffness and tumor progression, a variety of hydrogel scaffolds with dynamic changes in stiffness have been developed. These approaches, however, are not biocompatible at high temperature, strong irradiation, and acidic/basic pH, often lack reversibility (can only stiffen and not soften), and do not allow study on the same cell population longitudinally. In this work, we develop a dynamic 3D magnetic hydrogel whose matrix stiffness can be wirelessly and reversibly stiffened and softened multiple times with different rates of change using an external magnet. With this platform, we found that matrix stiffness increased tumor malignancy including denser cell organization, epithelial-to-mesenchymal transition and hypoxia. More interestingly, these malignant transformations could be halted or reversed with matrix softening (i.e., mechanical rescue), to potentiate drug efficacy attributing to reduced solid stress from matrix and downregulation of cell mechanotransductors including YAP1. We propose that our platform can be used to deepen understanding of the impact of matrix softening on cancer biology, an important but rarely studied phenomenon.

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Section: Discussionsupporting

confidence: 72%

Dynamic Magneto-Softening of 3D Hydrogel Reverses Malignant Transformation of Cancer Cells and Enhances Drug Efficacy

Shou

Teo

et al. 2023

ACS Nano

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“…Therefore, we also exploited the native biocompatibility of the NorHA hydrogel and the susceptibility of EAC PDOs to anti-cancer drugs to establish a novel in vivo model for targeted therapy studies. Whilst recent work has focused on understanding the role of engineered-ECM properties in tumor PDO (not EAC) resistance to therapy 23,[27][28][29] , we showed that our novel in vivo xenograft model allows for identification of matrix stiffness-dependent expression of transcription factors that can be exploited for targeted therapy in EAC. Therefore, this novel engineered organoid culture platform lays the foundation for application of therapeutics to disrupt contribution of ECM stiffness in EAC, and potentially, other cancers.…”

Section: Discussionmentioning

confidence: 96%

“…Indeed, HA-based hydrogels have been used by other groups to study tumor progression and resistance to therapy of other cancer types (e.g. colorectal and pancreatic adenocarcinomas) 28,29,37 .…”

Section: Engineered Hydrogel Supports Eac Pdo Developmentmentioning

confidence: 99%

“…Additionally, the engineered hydrogel served as a platform to identify potential therapeutic targets to disrupt the contribution of pro-tumorigenic increased matrix mechanics in EAC. Whereas previous work has established engineered hydrogels as tumor ECMs to investigate multicellular assembly and tumor invasion using cancer cell lines [24][25][26] , or study tumor PDO resistance to therapy 23,[27][28][29] , we are the first to analyze the novel contributions of ECM mechanical properties to EAC PDO growth, proliferation, and identification of matrix mechanics-mediated drivers of stem-like properties as therapeutic targets. Finally, the modular nature of the engineered hydrogel platform allows for potential adaption to the culture of 3D organoid models of other human cancers.…”

Section: Introductionmentioning

confidence: 99%

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Engineered hydrogel reveals contribution of matrix mechanics to esophageal adenocarcinoma 3D organoids and identify matrix-activated therapeutic targets

Cruz‐Acuña

Kariuki

Sugiura

et al. 2022

Preprint

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Increased extracellular matrix (ECM) stiffness has been implicated in esophageal adenocarcinoma (EAC) progression, metastasis, and resistance to therapy. However, the underlying pro-tumorigenic pathways are yet to be defined. Additional work is needed to develop physiologically relevant in vitro 3D culture models that better recapitulate the human tumor microenvironment and can be used to dissect the contributions of matrix stiffness to EAC pathogenesis. Here, we describe a modular, tumor ECM-mimetic hydrogel platform with tunable mechanical properties, defined presentation of cell-adhesive ligands, and protease-dependent degradation that supports robust in vitro growth and expansion of patient-derived EAC 3D organoids (EAC PDOs). Hydrogel mechanical properties control EAC PDO formation, growth, proliferation and activation of tumor-associated pathways that elicit stem-like properties in the cancer cells, as highlighted through in vitro and in vivo environments. We also demonstrate that the engineered hydrogel serves as a platform to identify potential therapeutic targets to disrupt the contribution of pro-tumorigenic increased matrix mechanics in EAC. Together, these studies show that an engineered PDO culture platform can be used to inform the development of therapeutics that target ECM stiffness in EAC.

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“…However, this workflow did not involve 3D biomatrix, which could limit the use of this technique for application, such as tumour organoids, where a bio-matrix is critical to maintain cell growth [39]. Further, it is known that stiffness of the culture biomatrix affects tumour cell phenotypes, such as chemosensitivity, therefore incorporating a matrix is likely critical for cancer culture platforms [2,40,41]. The use of a 3D matrix, as is typical in many engineered culture models however, significantly increases the complexity of image-based single cell analysis as these models are typically formed into a thick gel plug geometry with a curved meniscus surface profile.…”

Section: Introductionmentioning

confidence: 99%

An automation workflow for high-throughput manufacturing and analysis of scaffold-supported 3D tissue arrays

Cao

Cadavid

et al. 2022

Preprint

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The success rate of bringing novel cancer therapies to the clinic remains extremely low due to the lack of relevant pre-clinical culture models that capture the complexity of human tumours. Patient-derived organoids have emerged as a useful tool to model patient and tumour heterogeneity to begin addressing this need. Scaling these complex culture models while enabling stratified analysis of different cellular sub-populations remains a challenge, however. One strategy to enable higher throughput organoid cultures that also enables easy image-based analysis is the Scaffold-supported Platform for Organoid-based Tissues (SPOT) platform. SPOT allows the generation of flat, thin and dimensionally-defined microtissues in both 96- and 384-well plate footprints and is compatible with tumour organoid culture and downstream image-based readouts. SPOT manufacturing is currently a manual process however, limiting the use of SPOT to perform larger-scale screening. In this study, we integrate and optimize an automation approach to generate tumour-mimetic 3D engineered microtissues in SPOT using a liquid handler, and show comparable within-sample and between-sample variation as the standard manual manufacturing process. Furthermore, we develop a liquid handler-supported whole-cell extraction protocol and as a proof-of-value demonstration, we generate 3D complex tissues containing different proportions of tumour and stromal cells and perform single-cell-based end-point analysis to demonstrate the impact of co-culture on the tumour cell population specifically. We also demonstrate we can incorporate primary patient-derived organoids into the pipeline to capture patient-level tumour heterogeneity. We envision that this automated workflow integrated with 96/384-SPOT and multiple cell types and patient-derived organoid models will provide opportunities for future applications in high-throughput screening for novel personalized therapeutic targets. This pipeline also allows the user to assess dynamic cell responses using high-content longitudinal imaging or downstream single-cell-based analyses.

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Engineered extracellular matrices reveal stiffness-mediated chemoresistance in patient-derived pancreatic cancer organoids

Cited by 10 publications

References 72 publications

Dynamic Magneto-Softening of 3D Hydrogel Reverses Malignant Transformation of Cancer Cells and Enhances Drug Efficacy

Dynamic Magneto-Softening of 3D Hydrogel Reverses Malignant Transformation of Cancer Cells and Enhances Drug Efficacy

Engineered hydrogel reveals contribution of matrix mechanics to esophageal adenocarcinoma 3D organoids and identify matrix-activated therapeutic targets

An automation workflow for high-throughput manufacturing and analysis of scaffold-supported 3D tissue arrays

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