Putative critical quality attribute matrix identifies mesenchymal stromal cells with potent immunomodulatory and angiogenic “fitness” ranges in response to culture process parameters
Adipose-derived mesenchymal stromal cells (MSC(AT)) display immunomodulatory and angiogenic properties, but an improved understanding of quantitative critical quality attributes (CQAs) that inform basal MSC(AT) fitness ranges for immunomodulatory and/or angiogenic applications is urgently needed for...
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Frontiers Media S.A.
2022-11-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2022.972095/full |
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| author | Kevin P. Robb Kevin P. Robb Kevin P. Robb Julie Audet Rajiv Gandhi Rajiv Gandhi Sowmya Viswanathan Sowmya Viswanathan Sowmya Viswanathan Sowmya Viswanathan |
| author_facet | Kevin P. Robb Kevin P. Robb Kevin P. Robb Julie Audet Rajiv Gandhi Rajiv Gandhi Sowmya Viswanathan Sowmya Viswanathan Sowmya Viswanathan Sowmya Viswanathan |
| author_sort | Kevin P. Robb |
| collection | DOAJ |
| description | Adipose-derived mesenchymal stromal cells (MSC(AT)) display immunomodulatory and angiogenic properties, but an improved understanding of quantitative critical quality attributes (CQAs) that inform basal MSC(AT) fitness ranges for immunomodulatory and/or angiogenic applications is urgently needed for effective clinical translation. We constructed an in vitro matrix of multivariate readouts to identify putative CQAs that were sensitive enough to discriminate between specific critical processing parameters (CPPs) chosen for their ability to enhance MSC immunomodulatory and angiogenic potencies, with consideration for donor heterogeneity. We compared 3D aggregate culture conditions (3D normoxic, 3D-N) and 2D hypoxic (2D-H) culture as non-genetic CPP conditions that augment immunomodulatory and angiogenic fitness of MSC(AT). We measured multivariate panels of curated genes, soluble factors, and morphometric features for MSC(AT) cultured under varying CPP and licensing conditions, and we benchmarked these against two functional and therapeutically relevant anchor assays – in vitro monocyte/macrophage (MΦ) polarization and in vitro angiogenesis. Our results showed that varying CPP conditions was the primary driver of MSC(AT) immunomodulatory fitness; 3D-N conditions induced greater MSC(AT)-mediated MΦ polarization toward inflammation-resolving subtypes. In contrast, donor heterogeneity was the primary driver of MSC(AT) angiogenic fitness. Our analysis further revealed panels of putative CQAs with minimum and maximum values that consisted of twenty MSC(AT) characteristics that informed immunomodulatory fitness ranges, and ten MSC(AT) characteristics that informed angiogenic fitness ranges. Interestingly, many of the putative CQAs consisted of angiogenic genes or soluble factors that were inversely correlated with immunomodulatory functions (THBS1, CCN2, EDN1, PDGFA, VEGFA, EDIL3, ANGPT1, and ANG genes), and positively correlated to angiogenic functions (VEGF protein), respectively. We applied desirability analysis to empirically rank the putative CQAs for MSC(AT) under varying CPP conditions and donors to numerically identify the desirable CPP conditions or donors with maximal MSC(AT) immunomodulatory and/or angiogenic fitness. Taken together, our approach enabled combinatorial analysis of the matrix of multivariate readouts to provide putative quantitative CQAs that were sensitive to variations in select CPPs that enhance MSC immunomodulatory/angiogenic potency, and donor heterogeneity. These putative CQAs may be used to prospectively screen potent MSC(AT) donors or cell culture conditions to optimize for desired basal MSC(AT) immunomodulatory or angiogenic fitness. |
| first_indexed | 2024-04-11T15:53:47Z |
| format | Article |
| id | doaj.art-a55d030771e94d70873c06dce01014f7 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-04-11T15:53:47Z |
| publishDate | 2022-11-01 |
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| series | Frontiers in Immunology |
| spelling | doaj.art-a55d030771e94d70873c06dce01014f72022-12-22T04:15:13ZengFrontiers Media S.A.Frontiers in Immunology1664-32242022-11-011310.3389/fimmu.2022.972095972095Putative critical quality attribute matrix identifies mesenchymal stromal cells with potent immunomodulatory and angiogenic “fitness” ranges in response to culture process parametersKevin P. Robb0Kevin P. Robb1Kevin P. Robb2Julie Audet3Rajiv Gandhi4Rajiv Gandhi5Sowmya Viswanathan6Sowmya Viswanathan7Sowmya Viswanathan8Sowmya Viswanathan9Osteoarthritis Research Program, Division of Orthopedic Surgery, Schroeder Arthritis Institute, University Health Network, Toronto, ON, CanadaKrembil Research Institute, University Health Network, Toronto, ON, CanadaInstitute of Biomedical Engineering, University of Toronto, Toronto, ON, CanadaInstitute of Biomedical Engineering, University of Toronto, Toronto, ON, CanadaOsteoarthritis Research Program, Division of Orthopedic Surgery, Schroeder Arthritis Institute, University Health Network, Toronto, ON, CanadaDepartment of Surgery, Division of Orthopedic Surgery, University of Toronto, Toronto, ON, CanadaOsteoarthritis Research Program, Division of Orthopedic Surgery, Schroeder Arthritis Institute, University Health Network, Toronto, ON, CanadaKrembil Research Institute, University Health Network, Toronto, ON, CanadaInstitute of Biomedical Engineering, University of Toronto, Toronto, ON, CanadaDepartment of Medicine, Division of Hematology, University of Toronto, Toronto, ON, CanadaAdipose-derived mesenchymal stromal cells (MSC(AT)) display immunomodulatory and angiogenic properties, but an improved understanding of quantitative critical quality attributes (CQAs) that inform basal MSC(AT) fitness ranges for immunomodulatory and/or angiogenic applications is urgently needed for effective clinical translation. We constructed an in vitro matrix of multivariate readouts to identify putative CQAs that were sensitive enough to discriminate between specific critical processing parameters (CPPs) chosen for their ability to enhance MSC immunomodulatory and angiogenic potencies, with consideration for donor heterogeneity. We compared 3D aggregate culture conditions (3D normoxic, 3D-N) and 2D hypoxic (2D-H) culture as non-genetic CPP conditions that augment immunomodulatory and angiogenic fitness of MSC(AT). We measured multivariate panels of curated genes, soluble factors, and morphometric features for MSC(AT) cultured under varying CPP and licensing conditions, and we benchmarked these against two functional and therapeutically relevant anchor assays – in vitro monocyte/macrophage (MΦ) polarization and in vitro angiogenesis. Our results showed that varying CPP conditions was the primary driver of MSC(AT) immunomodulatory fitness; 3D-N conditions induced greater MSC(AT)-mediated MΦ polarization toward inflammation-resolving subtypes. In contrast, donor heterogeneity was the primary driver of MSC(AT) angiogenic fitness. Our analysis further revealed panels of putative CQAs with minimum and maximum values that consisted of twenty MSC(AT) characteristics that informed immunomodulatory fitness ranges, and ten MSC(AT) characteristics that informed angiogenic fitness ranges. Interestingly, many of the putative CQAs consisted of angiogenic genes or soluble factors that were inversely correlated with immunomodulatory functions (THBS1, CCN2, EDN1, PDGFA, VEGFA, EDIL3, ANGPT1, and ANG genes), and positively correlated to angiogenic functions (VEGF protein), respectively. We applied desirability analysis to empirically rank the putative CQAs for MSC(AT) under varying CPP conditions and donors to numerically identify the desirable CPP conditions or donors with maximal MSC(AT) immunomodulatory and/or angiogenic fitness. Taken together, our approach enabled combinatorial analysis of the matrix of multivariate readouts to provide putative quantitative CQAs that were sensitive to variations in select CPPs that enhance MSC immunomodulatory/angiogenic potency, and donor heterogeneity. These putative CQAs may be used to prospectively screen potent MSC(AT) donors or cell culture conditions to optimize for desired basal MSC(AT) immunomodulatory or angiogenic fitness.https://www.frontiersin.org/articles/10.3389/fimmu.2022.972095/fullmesenchymal stromal cellimmunomodulationangiogenesiscritical quality attributedonor heterogeneity3D suspension cultures |
| spellingShingle | Kevin P. Robb Kevin P. Robb Kevin P. Robb Julie Audet Rajiv Gandhi Rajiv Gandhi Sowmya Viswanathan Sowmya Viswanathan Sowmya Viswanathan Sowmya Viswanathan Putative critical quality attribute matrix identifies mesenchymal stromal cells with potent immunomodulatory and angiogenic “fitness” ranges in response to culture process parameters Frontiers in Immunology mesenchymal stromal cell immunomodulation angiogenesis critical quality attribute donor heterogeneity 3D suspension cultures |
| title | Putative critical quality attribute matrix identifies mesenchymal stromal cells with potent immunomodulatory and angiogenic “fitness” ranges in response to culture process parameters |
| title_full | Putative critical quality attribute matrix identifies mesenchymal stromal cells with potent immunomodulatory and angiogenic “fitness” ranges in response to culture process parameters |
| title_fullStr | Putative critical quality attribute matrix identifies mesenchymal stromal cells with potent immunomodulatory and angiogenic “fitness” ranges in response to culture process parameters |
| title_full_unstemmed | Putative critical quality attribute matrix identifies mesenchymal stromal cells with potent immunomodulatory and angiogenic “fitness” ranges in response to culture process parameters |
| title_short | Putative critical quality attribute matrix identifies mesenchymal stromal cells with potent immunomodulatory and angiogenic “fitness” ranges in response to culture process parameters |
| title_sort | putative critical quality attribute matrix identifies mesenchymal stromal cells with potent immunomodulatory and angiogenic fitness ranges in response to culture process parameters |
| topic | mesenchymal stromal cell immunomodulation angiogenesis critical quality attribute donor heterogeneity 3D suspension cultures |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2022.972095/full |
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