Tunable hybrid hydrogels with multicellular spheroids for modeling desmoplastic pancreatic cancer

Menekse Ermis; Natashya Falcone; Natan Roberto de Barros; Marvin Mecwan; Reihaneh Haghniaz; Auveen Choroomi; Mahsa Monirizad; Yeji Lee; Jihyeon Song; Hyun Jong Cho; Yangzhi Zhu; Heemin Kang; Mehmet R. Dokmeci; Ali Khademhosseini; Junmin Lee; Han Jun Kim

doi:10.1016/j.bioactmat.2023.02.005

Tunable hybrid hydrogels with multicellular spheroids for modeling desmoplastic pancreatic cancer

Menekse Ermis, Natashya Falcone, Natan Roberto de Barros, Marvin Mecwan, Reihaneh Haghniaz, Auveen Choroomi, Mahsa Monirizad, Yeji Lee, Jihyeon Song, Hyun Jong Cho, Yangzhi Zhu, Heemin Kang, Mehmet R. Dokmeci, Ali Khademhosseini, Junmin Lee, Han Jun Kim

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25 Citations (Scopus)

Abstract

The tumor microenvironment consists of diverse, complex etiological factors. The matrix component of pancreatic ductal adenocarcinoma (PDAC) plays an important role not only in physical properties such as tissue rigidity but also in cancer progression and therapeutic responsiveness. Although significant efforts have been made to model desmoplastic PDAC, existing models could not fully recapitulate the etiology to mimic and understand the progression of PDAC. Here, two major components in desmoplastic pancreatic matrices, hyaluronic acid- and gelatin-based hydrogels, are engineered to provide matrices for tumor spheroids composed of PDAC and cancer-associated fibroblasts (CAF). Shape analysis profiles reveals that incorporating CAF contributes to a more compact tissue formation. Higher expression levels of markers associated with proliferation, epithelial to mesenchymal transition, mechanotransduction, and progression are observed for cancer-CAF spheroids cultured in hyper desmoplastic matrix-mimicking hydrogels, while the trend can be observed when those are cultured in desmoplastic matrix-mimicking hydrogels with the presence of transforming growth factor-β1 (TGF-β1). The proposed multicellular pancreatic tumor model, in combination with proper mechanical properties and TGF-β1 supplement, makes strides in developing advanced pancreatic models for resembling and monitoring the progression of pancreatic tumors, which could be potentially applicable for realizing personalized medicine and drug testing applications.

Original language	English
Pages (from-to)	360-373
Number of pages	14
Journal	Bioactive Materials
Volume	25
DOIs	https://doi.org/10.1016/j.bioactmat.2023.02.005
Publication status	Published - 2023 Jul

Bibliographical note

Funding Information:
In tumor progression, TMEs provide a unique physical and biochemical niche for tumor growth [46]. To implement a TME composed of complex and diverse cellular/ECM components, in this study, we tuned the cellular, ECM and tissue stiffness aspects of the tumor niche and analyzed the concomitant biological response of the microtumor model. To this end, tumor/CAF spheroids were embedded into a tunable HAMA/GelMA hybrid hydrogel, and the degree of crosslinking was adjusted to simulate normal, desmoplastic, and hyperdesmoplastic matrices. Live/dead viability assays and metabolic activity assays were performed to assess viability within three different stiffnesses of HAMA/GelMA hydrogels (3, 15, 36 kPa) over time (Fig. 3A). In both live/dead assay and PrestoBlue metabolic activity assay, it was confirmed that HAMA/GelMA hydrogel under experimental conditions did not adversely affect cell viability. In all conditions, viability was observed above 95% at day 1 and cell proliferation increased over time. Interestingly, the spheroids composed of mixture of cancer cells and CAFs showed no significant difference in viability on day 7, whereas the spheroids composed only of cancer cells showed differences in viability according to the stiffness of the HAMA/GelMA hybrid hydrogel. In particular, the viability increased as the stiffness of the HAMA/GelMA hybrid hydrogel decreased (Fig. 3A, Fig. S6). These results suggest that both the presence of CAF and the stiffness of the embedding gel can influence the proliferation of cellular components within the microtumor model. Bauer et al. demonstrated that CAFs are mechanosensitive, as the substrate stiffness increases up to a critical value (20–40 kPa). In this study, CAFs responded to substrate stiffness by applying more force to the hydrogels as well as secreting cytokines that activate the TGF-β pathway [47]. Calvo et al. also tested CAFs with different substrate stiffness that as the substrate stiffness increased, the number of YAP (a mechanosensitive transcription factor that translocated to the nucleus and activated genes regulating matrix regulation)-positive nuclei increased, demonstrating the mechanosensitive nature of CAF in cancer [48]. This study supports our finding that the group with CAF can adapt to the environment, maintaining relatively constant viability, even with increased stiffness of the TME, such as desmoplastic and hyperdesmoplastic conditions, compared to the group without CAF.Dr. M. Ermis and Dr. N. Falcone contributed equally to this work. The authors gratefully acknowledge the funding by the National Institutes of Health (HL140951, HL137193, CA257558, DK130566). Dr. M. Ermis acknowledges The Scientific and Technological Research Council of Turkiye for 2219-International Postdoctoral Research Fellowship Program.

Funding Information:
Dr. M. Ermis and Dr. N. Falcone contributed equally to this work. The authors gratefully acknowledge the funding by the National Institutes of Health ( HL140951 , HL137193 , CA257558 , DK130566 ). Dr. M. Ermis acknowledges The Scientific and Technological Research Council of Turkiye for 2219-International Postdoctoral Research Fellowship Program.

Publisher Copyright:
© 2023 The Authors

Keywords

Desmoplasia
Extracellular matrix
Fibrosis
Pancreatic cancer
Tumor microenvironment

ASJC Scopus subject areas

Biotechnology
Biomaterials
Biomedical Engineering

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.bioactmat.2023.02.005

Cite this

Ermis, M., Falcone, N., Roberto de Barros, N., Mecwan, M., Haghniaz, R., Choroomi, A., Monirizad, M., Lee, Y., Song, J., Cho, H. J., Zhu, Y., Kang, H., Dokmeci, M. R., Khademhosseini, A., Lee, J., & Kim, H. J. (2023). Tunable hybrid hydrogels with multicellular spheroids for modeling desmoplastic pancreatic cancer. Bioactive Materials, 25, 360-373. https://doi.org/10.1016/j.bioactmat.2023.02.005

Ermis, M, Falcone, N, Roberto de Barros, N, Mecwan, M, Haghniaz, R, Choroomi, A, Monirizad, M, Lee, Y, Song, J, Cho, HJ, Zhu, Y, Kang, H, Dokmeci, MR, Khademhosseini, A, Lee, J & Kim, HJ 2023, 'Tunable hybrid hydrogels with multicellular spheroids for modeling desmoplastic pancreatic cancer', Bioactive Materials, vol. 25, pp. 360-373. https://doi.org/10.1016/j.bioactmat.2023.02.005

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title = "Tunable hybrid hydrogels with multicellular spheroids for modeling desmoplastic pancreatic cancer",

abstract = "The tumor microenvironment consists of diverse, complex etiological factors. The matrix component of pancreatic ductal adenocarcinoma (PDAC) plays an important role not only in physical properties such as tissue rigidity but also in cancer progression and therapeutic responsiveness. Although significant efforts have been made to model desmoplastic PDAC, existing models could not fully recapitulate the etiology to mimic and understand the progression of PDAC. Here, two major components in desmoplastic pancreatic matrices, hyaluronic acid- and gelatin-based hydrogels, are engineered to provide matrices for tumor spheroids composed of PDAC and cancer-associated fibroblasts (CAF). Shape analysis profiles reveals that incorporating CAF contributes to a more compact tissue formation. Higher expression levels of markers associated with proliferation, epithelial to mesenchymal transition, mechanotransduction, and progression are observed for cancer-CAF spheroids cultured in hyper desmoplastic matrix-mimicking hydrogels, while the trend can be observed when those are cultured in desmoplastic matrix-mimicking hydrogels with the presence of transforming growth factor-β1 (TGF-β1). The proposed multicellular pancreatic tumor model, in combination with proper mechanical properties and TGF-β1 supplement, makes strides in developing advanced pancreatic models for resembling and monitoring the progression of pancreatic tumors, which could be potentially applicable for realizing personalized medicine and drug testing applications.",

keywords = "Desmoplasia, Extracellular matrix, Fibrosis, Pancreatic cancer, Tumor microenvironment",

author = "Menekse Ermis and Natashya Falcone and {Roberto de Barros}, Natan and Marvin Mecwan and Reihaneh Haghniaz and Auveen Choroomi and Mahsa Monirizad and Yeji Lee and Jihyeon Song and Cho, {Hyun Jong} and Yangzhi Zhu and Heemin Kang and Dokmeci, {Mehmet R.} and Ali Khademhosseini and Junmin Lee and Kim, {Han Jun}",

note = "Funding Information: In tumor progression, TMEs provide a unique physical and biochemical niche for tumor growth [46]. To implement a TME composed of complex and diverse cellular/ECM components, in this study, we tuned the cellular, ECM and tissue stiffness aspects of the tumor niche and analyzed the concomitant biological response of the microtumor model. To this end, tumor/CAF spheroids were embedded into a tunable HAMA/GelMA hybrid hydrogel, and the degree of crosslinking was adjusted to simulate normal, desmoplastic, and hyperdesmoplastic matrices. Live/dead viability assays and metabolic activity assays were performed to assess viability within three different stiffnesses of HAMA/GelMA hydrogels (3, 15, 36 kPa) over time (Fig. 3A). In both live/dead assay and PrestoBlue metabolic activity assay, it was confirmed that HAMA/GelMA hydrogel under experimental conditions did not adversely affect cell viability. In all conditions, viability was observed above 95% at day 1 and cell proliferation increased over time. Interestingly, the spheroids composed of mixture of cancer cells and CAFs showed no significant difference in viability on day 7, whereas the spheroids composed only of cancer cells showed differences in viability according to the stiffness of the HAMA/GelMA hybrid hydrogel. In particular, the viability increased as the stiffness of the HAMA/GelMA hybrid hydrogel decreased (Fig. 3A, Fig. S6). These results suggest that both the presence of CAF and the stiffness of the embedding gel can influence the proliferation of cellular components within the microtumor model. Bauer et al. demonstrated that CAFs are mechanosensitive, as the substrate stiffness increases up to a critical value (20–40 kPa). In this study, CAFs responded to substrate stiffness by applying more force to the hydrogels as well as secreting cytokines that activate the TGF-β pathway [47]. Calvo et al. also tested CAFs with different substrate stiffness that as the substrate stiffness increased, the number of YAP (a mechanosensitive transcription factor that translocated to the nucleus and activated genes regulating matrix regulation)-positive nuclei increased, demonstrating the mechanosensitive nature of CAF in cancer [48]. This study supports our finding that the group with CAF can adapt to the environment, maintaining relatively constant viability, even with increased stiffness of the TME, such as desmoplastic and hyperdesmoplastic conditions, compared to the group without CAF.Dr. M. Ermis and Dr. N. Falcone contributed equally to this work. The authors gratefully acknowledge the funding by the National Institutes of Health (HL140951, HL137193, CA257558, DK130566). Dr. M. Ermis acknowledges The Scientific and Technological Research Council of Turkiye for 2219-International Postdoctoral Research Fellowship Program. Funding Information: Dr. M. Ermis and Dr. N. Falcone contributed equally to this work. The authors gratefully acknowledge the funding by the National Institutes of Health ( HL140951 , HL137193 , CA257558 , DK130566 ). Dr. M. Ermis acknowledges The Scientific and Technological Research Council of Turkiye for 2219-International Postdoctoral Research Fellowship Program. Publisher Copyright: {\textcopyright} 2023 The Authors",

year = "2023",

month = jul,

doi = "10.1016/j.bioactmat.2023.02.005",

language = "English",

volume = "25",

pages = "360--373",

journal = "Bioactive Materials",

issn = "2452-199X",

publisher = "KeAi Communications Co",

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TY - JOUR

T1 - Tunable hybrid hydrogels with multicellular spheroids for modeling desmoplastic pancreatic cancer

AU - Ermis, Menekse

AU - Falcone, Natashya

AU - Roberto de Barros, Natan

AU - Mecwan, Marvin

AU - Haghniaz, Reihaneh

AU - Choroomi, Auveen

AU - Monirizad, Mahsa

AU - Lee, Yeji

AU - Song, Jihyeon

AU - Cho, Hyun Jong

AU - Zhu, Yangzhi

AU - Kang, Heemin

AU - Dokmeci, Mehmet R.

AU - Khademhosseini, Ali

AU - Lee, Junmin

AU - Kim, Han Jun

N1 - Funding Information: In tumor progression, TMEs provide a unique physical and biochemical niche for tumor growth [46]. To implement a TME composed of complex and diverse cellular/ECM components, in this study, we tuned the cellular, ECM and tissue stiffness aspects of the tumor niche and analyzed the concomitant biological response of the microtumor model. To this end, tumor/CAF spheroids were embedded into a tunable HAMA/GelMA hybrid hydrogel, and the degree of crosslinking was adjusted to simulate normal, desmoplastic, and hyperdesmoplastic matrices. Live/dead viability assays and metabolic activity assays were performed to assess viability within three different stiffnesses of HAMA/GelMA hydrogels (3, 15, 36 kPa) over time (Fig. 3A). In both live/dead assay and PrestoBlue metabolic activity assay, it was confirmed that HAMA/GelMA hydrogel under experimental conditions did not adversely affect cell viability. In all conditions, viability was observed above 95% at day 1 and cell proliferation increased over time. Interestingly, the spheroids composed of mixture of cancer cells and CAFs showed no significant difference in viability on day 7, whereas the spheroids composed only of cancer cells showed differences in viability according to the stiffness of the HAMA/GelMA hybrid hydrogel. In particular, the viability increased as the stiffness of the HAMA/GelMA hybrid hydrogel decreased (Fig. 3A, Fig. S6). These results suggest that both the presence of CAF and the stiffness of the embedding gel can influence the proliferation of cellular components within the microtumor model. Bauer et al. demonstrated that CAFs are mechanosensitive, as the substrate stiffness increases up to a critical value (20–40 kPa). In this study, CAFs responded to substrate stiffness by applying more force to the hydrogels as well as secreting cytokines that activate the TGF-β pathway [47]. Calvo et al. also tested CAFs with different substrate stiffness that as the substrate stiffness increased, the number of YAP (a mechanosensitive transcription factor that translocated to the nucleus and activated genes regulating matrix regulation)-positive nuclei increased, demonstrating the mechanosensitive nature of CAF in cancer [48]. This study supports our finding that the group with CAF can adapt to the environment, maintaining relatively constant viability, even with increased stiffness of the TME, such as desmoplastic and hyperdesmoplastic conditions, compared to the group without CAF.Dr. M. Ermis and Dr. N. Falcone contributed equally to this work. The authors gratefully acknowledge the funding by the National Institutes of Health (HL140951, HL137193, CA257558, DK130566). Dr. M. Ermis acknowledges The Scientific and Technological Research Council of Turkiye for 2219-International Postdoctoral Research Fellowship Program. Funding Information: Dr. M. Ermis and Dr. N. Falcone contributed equally to this work. The authors gratefully acknowledge the funding by the National Institutes of Health ( HL140951 , HL137193 , CA257558 , DK130566 ). Dr. M. Ermis acknowledges The Scientific and Technological Research Council of Turkiye for 2219-International Postdoctoral Research Fellowship Program. Publisher Copyright: © 2023 The Authors

PY - 2023/7

Y1 - 2023/7

N2 - The tumor microenvironment consists of diverse, complex etiological factors. The matrix component of pancreatic ductal adenocarcinoma (PDAC) plays an important role not only in physical properties such as tissue rigidity but also in cancer progression and therapeutic responsiveness. Although significant efforts have been made to model desmoplastic PDAC, existing models could not fully recapitulate the etiology to mimic and understand the progression of PDAC. Here, two major components in desmoplastic pancreatic matrices, hyaluronic acid- and gelatin-based hydrogels, are engineered to provide matrices for tumor spheroids composed of PDAC and cancer-associated fibroblasts (CAF). Shape analysis profiles reveals that incorporating CAF contributes to a more compact tissue formation. Higher expression levels of markers associated with proliferation, epithelial to mesenchymal transition, mechanotransduction, and progression are observed for cancer-CAF spheroids cultured in hyper desmoplastic matrix-mimicking hydrogels, while the trend can be observed when those are cultured in desmoplastic matrix-mimicking hydrogels with the presence of transforming growth factor-β1 (TGF-β1). The proposed multicellular pancreatic tumor model, in combination with proper mechanical properties and TGF-β1 supplement, makes strides in developing advanced pancreatic models for resembling and monitoring the progression of pancreatic tumors, which could be potentially applicable for realizing personalized medicine and drug testing applications.

AB - The tumor microenvironment consists of diverse, complex etiological factors. The matrix component of pancreatic ductal adenocarcinoma (PDAC) plays an important role not only in physical properties such as tissue rigidity but also in cancer progression and therapeutic responsiveness. Although significant efforts have been made to model desmoplastic PDAC, existing models could not fully recapitulate the etiology to mimic and understand the progression of PDAC. Here, two major components in desmoplastic pancreatic matrices, hyaluronic acid- and gelatin-based hydrogels, are engineered to provide matrices for tumor spheroids composed of PDAC and cancer-associated fibroblasts (CAF). Shape analysis profiles reveals that incorporating CAF contributes to a more compact tissue formation. Higher expression levels of markers associated with proliferation, epithelial to mesenchymal transition, mechanotransduction, and progression are observed for cancer-CAF spheroids cultured in hyper desmoplastic matrix-mimicking hydrogels, while the trend can be observed when those are cultured in desmoplastic matrix-mimicking hydrogels with the presence of transforming growth factor-β1 (TGF-β1). The proposed multicellular pancreatic tumor model, in combination with proper mechanical properties and TGF-β1 supplement, makes strides in developing advanced pancreatic models for resembling and monitoring the progression of pancreatic tumors, which could be potentially applicable for realizing personalized medicine and drug testing applications.

KW - Desmoplasia

KW - Extracellular matrix

KW - Fibrosis

KW - Pancreatic cancer

KW - Tumor microenvironment

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DO - 10.1016/j.bioactmat.2023.02.005

M3 - Article

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VL - 25

SP - 360

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JO - Bioactive Materials

JF - Bioactive Materials

ER -

Tunable hybrid hydrogels with multicellular spheroids for modeling desmoplastic pancreatic cancer

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

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