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US-20260124135-A1 - IMPLANTABLE SCAFFOLDS AND METHODS OF USE

US20260124135A1US 20260124135 A1US20260124135 A1US 20260124135A1US-20260124135-A1

Abstract

The present disclosure provides compositions, systems, and methods related to cellular transduction. In particular, the present disclosure provides compositions, systems, and methods pertaining to implantable macroporous scaffolds that facilitate rapid and highly efficient cellular transduction.

Inventors

  • Yevgeny Brudno
  • Madelyn VANBLUNK
  • Sharda Pandit
  • Pritha AGARWALLA
  • Gianpietro Dotti

Assignees

  • NORTH CAROLINA STATE UNIVERSITY
  • THE UNIVERSITY OF NORTH CAROLINA AT CHAPEL HILL

Dates

Publication Date
20260507
Application Date
20231013

Claims (20)

  1. 1 . An implantable macroporous scaffold comprising: a crosslinked biopolymer matrix comprising an average pore size ranging from about 10 μm to about 500 μm, and a stiffness ranging from about 1 kPa to about 1000 kPa, wherein the stiffness of the matrix is compatible with the stiffness of a target tissue; and a composition comprising a plurality of cells and a transduction agent; wherein the scaffold facilitates transduction of the plurality of cells with the transduction agent.
  2. 2 . The macroporous scaffold of claim 1 , wherein the biopolymer matrix comprises at least one of alginate, Hyaluronic acid, collagen, fibrin, Poly Lactic-co-Glycolic Acid (PLGA), Polycaprolactone (PCL), gelatin, Polyethylene glycol (PEG), chitosan, cellulose, polyglutamic acid, fibrin, silk, agarose, dextran, polyacrylamide, polyvinyl alcohol, Poly(N-isopropylacrylamide), Poly(2-hydroxyethyl methacrylate), polyurethane, polyethyleneimine, Poly(methyl methacrylate, Poly(2-oxazoline), Polyphosphazenes, and any composites, derivatives, or combinations thereof.
  3. 3 . The macroporous scaffold of claim 1 or claim 2 , wherein the biopolymer matrix comprises alginate having a molecular weight from about 1 kDa to about 500 kDa.
  4. 4 . The macroporous scaffold of any one of claims 1 to 3 , wherein the biopolymer matrix comprises alginate having a G/M ratio from about 0.5 to about 5.0.
  5. 5 . The macroporous scaffold of any one of claims 1 to 4 , wherein the biopolymer matrix comprises alginate at a concentration ranging from about 0.1% to about 5.0% (w/v).
  6. 6 . The macroporous scaffold of any one of claims 1 to 4 , wherein the biopolymer matrix comprises alginate at a concentration ranging from about 0.5% to about 2.0% (w/v).
  7. 7 . The macroporous scaffold of any one of claims 1 to 6 , wherein the biopolymer matrix comprises alginate at a concentration ranging from about 0.5% to about 1.5% (w/v).
  8. 8 . The macroporous scaffold of any one of claims 1 to 6 , wherein the biopolymer matrix comprises alginate at a concentration ranging from about 0.5% to about 1.0% (w/v).
  9. 9 . The macroporous scaffold of any one of claims 1 to 6 , wherein the biopolymer matrix comprises alginate at a concentration ranging from about 1.0% to about 2.0% (w/v).
  10. 10 . The macroporous scaffold of any one of claims 1 to 6 , wherein the biopolymer matrix comprises alginate at a concentration ranging from about 1.0% to about 1.5% (w/v).
  11. 11 . The macroporous scaffold of any one of claims 1 to 6 , wherein the biopolymer matrix comprises alginate at a concentration ranging from about 1.5% to about 2.0% (w/v).
  12. 12 . The macroporous scaffold of any one of claims 1 to 11 , wherein the biopolymer matrix comprises calcium alginate having a calcium concentration ranging from about 0.1% to about 1.0% (w/v).
  13. 13 . The macroporous scaffold of any one of claims 1 to 11 , wherein the biopolymer matrix comprises calcium alginate having a calcium concentration ranging from about 0.1% to about 0.3% (w/v).
  14. 14 . The macroporous scaffold of any one of claims 1 to 11 , wherein the biopolymer matrix comprises calcium alginate having a calcium concentration ranging from about 0.1% to about 0.2% (w/v).
  15. 15 . The macroporous scaffold of any one of claims 1 to 11 , wherein the biopolymer matrix comprises calcium alginate having a calcium concentration ranging from about 0.2% to about 0.3% (w/v).
  16. 16 . The macroporous scaffold of any one of claims 1 to 15 , wherein the biopolymer matrix is generated at a temperature ranging from about 0° C. to about −80° C.
  17. 17 . The macroporous scaffold of any one of claims 1 to 16 , wherein the biopolymer matrix exhibits a stiffness that is from about ±25%, about ±50%, about ±75%, about ±100%, about ±125%, about ±150%, about ±175%, about ±200%, about ±225%, or about ±250% of the stiffness of the target tissue.
  18. 18 . The macroporous scaffold of any one of claims 1 to 17 , wherein the scaffold comprises at least one biological agent.
  19. 19 . The macroporous scaffold of claim 18 , wherein the at least one biological agent is a small molecule.
  20. 20 . The macroporous scaffold of claim 19 , wherein the small molecule is selected from the group consisting of a TLR agonist, a checkpoint inhibitor, an IDO inhibitor, a MEK inhibitor, an HDAC inhibitor, a PI3K inhibitor, an immunomodulatory drug, a JAK kinase inhibitor, and an mTOR inhibitor.

Description

CROSS REFERENCE TO RELATED APPLICATIONS This application claims priority to and the benefit of U.S. Provisional Patent Application No. 63/415,806 filed Oct. 13, 2022, and U.S. Provisional Patent Application No. 63/441,409 filed Jan. 26, 2023, both of which are incorporated herein by reference in their entireties and for all purposes. GOVERNMENT SUPPORT This invention was made with government support under grant number CA260223 awarded by the National Institutes of Health. The government has certain rights in the invention. FIELD The present disclosure provides compositions, systems, and methods related to cellular transduction. In particular, the present disclosure provides compositions, systems, and methods pertaining to implantable macroporous scaffolds that facilitate rapid and highly efficient cellular transduction. BACKGROUND Chimeric Antigen Receptor (CAR) T cell therapy has had revolutionary clinical success in hematological cancers and demonstrated potential against a wide array of cancer types. Unfortunately, CAR T cell therapy has not to date had the same impact in solid tumors. This challenge stems from several limitations of current CAR T cell products, including extensive manufacturing procedures, therapeutic toxicity, limited in vivo persistence, and inability to achieve long term therapeutic efficacy. The costly and labor-intensive CAR T cell manufacturing presents several challenges for the success of these therapies across various hematological and solid malignancies. Current CAR T cell manufacturing requires extensive infrastructure and takes multiple weeks for vein-to-vein procedures, delaying the initiation of the treatment in patients with already advancing disease. These long manufacturing times (>2 weeks) and high treatment costs (˜$350K) limit widespread use of this therapy. Approaches to simplify this process include automated manufacturing, use of allogeneic products, and rapid CAR T cell manufacturing eliminating one or more CAR T cells generation steps. However, these approaches are limited by poor quality products, life-threatening toxicities, low persistence, and precocious cell differentiation and exhaustion. Generating and expanding tumor-specific CAR T cells in vivo could mitigate some of these problems by reducing manufacturing time, treatment cost, and producing long lasting highly efficacious cell products. In vivo expansion and long-term persistence of infused CAR T cells is critical to provide antitumor function and prevent tumor relapse. The importance of T cell persistence and long-term function is particularly acute in solid tumors, where physiological and immunological barriers such as hypoxia and the immunosuppressive tumor microenvironment alter CAR T cell metabolism and promote CAR T cell exhaustion leading to limited therapeutic outcomes. Recent work suggests that less differentiated CAR T cells could increase cell engraftment and persistence, leading to improved outcomes. Strategies to produce less differentiated CAR T cell phenotypes show promise and include an initial selection of naïve cells, optimization of in vitro culture time and cytokine regiments as well as addition of small molecule T cell regulators. Despite extensive studies, initial selection of naïve population or maintaining naïve population during clinical-grade manufacturing of CAR T cells is associated with multiple technical difficulties. A simple, scalable, and tunable CAR T cell manufacturing platform to generate CAR T cells with less differentiated phenotypes would improve long-term persistence and therapeutic efficacy. Biomaterials could overcome many of the obstacles to widespread, safe, and efficacious CAR T cell therapy for solid tumors by providing a nurturing niche to optimize cell development. Recent efforts have used biomaterials to improve specific, individual steps of CAR T cell manufacturing, including in vitro and in vivo T cell isolation, T cell activation, genetic modification, expansion, and delivery. In addition, biomaterials were used to control T cell proliferation and differentiation to produce robust antitumor effects. Finally, biomaterials enable sustained release of CAR T cells, which could improve the safety and toxicity profile of CAR T cell therapy. Recent work suggests that splitting CAR T cell administration into multiple, smaller doses can lower the toxic profile and expand the therapeutic window of this therapy. Sustained release enables an improved multi-dose schedule creating an avenue for significant CAR T cell doses without an initial toxic burst of effector function. SUMMARY Embodiments of the present disclosure include an implantable macroporous scaffold comprising a crosslinked biopolymer matrix comprising an average pore size ranging from about 10 μm to about 500 μm, and a stiffness ranging from about 1 kPa to about 1000 kPa, wherein the stiffness of the matrix is compatible with the stiffness of a target tissue; and a composition comprising a plurality