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Experimental and Computational Models of Transport of Galectin-3 Through Glycosylated Matrix

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Abstract

Altered extracellular matrix (ECM) production is a hallmark of many fibroproliferative diseases, including certain cancers. The high incidence of glycan-rich components within altered ECM makes the use of glycan-binding proteins such as Galectin-3 (G3) a promising therapeutic strategy. The complexity of ECM as a rich 3D network of proteins with varied glycosylation states makes it challenging to determine the retention of glycan-binding proteins in altered ECM environments. Computational models capable of predicting the transport of glycan-binding proteins in altered ECM can benefit the design and testing of such proteins and associated novel therapeutic strategies. However, such computational models require many kinetic parameters that cannot be estimated from traditional 2D pharmacokinetic assays. To validate transport properties of G3 in 3D ECM constructs, we developed a species transport model that includes diffusion and matrix-binding components to predict retention of G3 fusion proteins in glycan-rich ECM. By iteratively comparing our computational model to experimental results, we are able to determine a reasonable range of parameters for a robust computational model of G3 transport. We anticipate this overall approach to building a data-driven model is translatable to other ECM-targeting therapeutic strategies.

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Abbreviations

ASF:

Asialofetuin

k f :

Association constant

k f_ColIV :

Association constant Collagen IV

k f_Lam :

Association constant Laminin

N :

Available binding sites per molecule

N ColIV :

Available binding sites per molecule of CollagenIV

N Lam :

Available binding sites per molecule of Laminin

Col I:

Collagen Type I

Col IV:

Collagen Type IV

k r :

Dissociation constant

D g :

Effective diffusivity in gel

K D :

Equilibrium dissociation constant

K DColIV :

Equilibrium dissociation constant Collagen IV

C eq :

Equilibrium media concentration of GFP-G3

ϕ :

Fluid volume fraction

G3:

Galectin-3

C 0 :

Initial concentration of GFP-G3 in the gel per unit volume of gel

Lam:

Laminin

C gp :

Molar concentration of glycoprotein

GFP-G3:

Superfolder green fluorescence protein fused to Galectin-3

R free :

Volume average concentration of free ECM receptors for GFP-G3

C bound :

Volume average concentration of GFP-G3 bound to gel

R 0 :

Volume average concentration of the total binding sites in gel

V g :

Volume of gel

V m :

Volume of media

β-lac:

β-Lactose

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Acknowledgments

We thank Dr. Natalie Fredette and John Sansalone for their technical assistance preparing initial Galectin 3 experiments. We also thank Edwin Rajeev for his assistance developing initial computational transport models.

Author Contributions

CSS conceived of the project and, in collaboration with MS and GAH, oversaw overall direction, planning experiments, data analysis, and writing and editing the manuscript. JPLL contributed to the experimental design, conducted the experimental measurements, analyzed the data, and contributed to the manuscript’s writing and figure design. SAF contributed to the fabrication of G3 fusion protein and the development of protocols for several experiments. CR performed the computational simulations for the sensitivity analysis and contributed to data analysis. MS derived the computational models, conducted computational simulations, analyzed data, and contributed to writing and editing the manuscript. GAH contributed to the development of protocols for several experiments, provided guidance, and contributed to the interpretation of the results. All authors provided critical feedback and helped shape the research, analysis, and manuscript.

Funding

This work was partially funded by the National Institutes of Health [R03 EB019684 to G.A.H]; National Institutes of Health-National Center for Advancing Translational Sciences under the University of Florida Clinical and Translational Award [TL1TR001428 to S.A.F]; and the UF Research Opportunity Seed Fund [to M.S.].

Conflict of interest

Dr.Gregory A. Hudalla is a founder, scientific advisory member, and stockholder of Anchor Biologics, Inc. which is developing technologies related to galectin fusion proteins.

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Authors and Affiliations

  1. J. Crayton Pruitt Family Department of Biomedical Engineering, Herbert Wertheim College of Engineering, University of Florida, Gainesville, FL, 32611, USA

    Janny Piñeiro-Llanes, Shaheen A. Farhadi, Gregory A. Hudalla, Malisa Sarntinoranont & Chelsey S. Simmons

  2. Department of Mechanical and Aerospace Engineering, Herbert Wertheim College of Engineering, University of Florida, Gainesville, FL, 32611, USA

    Camille D. Rodriguez, Malisa Sarntinoranont & Chelsey S. Simmons

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  1. Janny Piñeiro-Llanes
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  2. Camille D. Rodriguez
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  3. Shaheen A. Farhadi
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  4. Gregory A. Hudalla
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  6. Chelsey S. Simmons
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Correspondence to Chelsey S. Simmons.

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Associate Editor Shayn Peirce-Cottler oversaw the review of this article.

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Piñeiro-Llanes, J., Rodriguez, C.D., Farhadi, S.A. et al. Experimental and Computational Models of Transport of Galectin-3 Through Glycosylated Matrix. Ann Biomed Eng 50, 703–715 (2022). https://doi.org/10.1007/s10439-022-02949-6

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