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JP-7854806-B2 - Patch transplantation of stem cells/precursors into solid organs

JP7854806B2JP 7854806 B2JP7854806 B2JP 7854806B2JP-7854806-B2

Inventors

  • リード，ローラ エム．
  • ザン，ウェンチェン
  • ワウティア，エリアン

Assignees

  • ザ ユニバーシティ オブ ノース カロライナ アット チャペル ヒル

Dates

Publication Date
20260507
Application Date
20200522
Priority Date
20190524

Claims (10)

  1. A patch graft comprising a mixture of epithelial cells and mesenchymal cells for use in a method to induce, restore, increase, or improve the function of a target disease, disorder, or dysfunctional liver suffering from type 1 tyrosinemia, The method includes contacting the diseased, impaired, or dysfunctional liver with a patch graft containing a mixture of epithelial cells and mesenchymal cells, such that the introduction, restoration, increase, or improvement of the functionality of the diseased, impaired, or dysfunctional liver is manifested by measuring a decrease in the levels of tyrosine or alpha-fetoprotein measured in a biological sample obtained from the subject, under conditions that promote the engraftment of epithelial cells and mesenchymal cells. A patch graft containing epithelial cells, including bile dendritic stem cells (BTSCs), and mesenchymal cells, including early lineage mesenchymal cells (ELSMCs).
  2. The patch graft according to claim 1, wherein the exposed surface of the patch graft includes a covering that inhibits adhesion of the patch graft to nearby organs and tissues.
  3. A patch graft according to any one of claims 1 to 2, comprising one or more biomaterial layers, wherein the biomaterial layers are at least a) A first inner layer for contacting solid organs, exhibiting first viscoelastic properties, incorporating a mixture of epithelial cells and mesenchymal cells, supporting the ability of epithelial and mesenchymal cells to produce secretory matrix metalloproteinases (MMPs), and promoting the viability and immaturity of the aforementioned epithelial and mesenchymal cells. Includes, Depending on the circumstances, b ) A backing that provides a barrier to cells migrating in directions other than solid organs, and which exhibits a second viscoelastic property , and/or c ) A third outer layer of covering or material that exhibits third viscoelastic properties, which minimizes adhesion of the patch graft to the inner wall and/or inner surface of the body cavity, including organs, in the vicinity of the patch graft . It may include , A patch graft in which the aforementioned viscoelastic properties are determined by measuring the fluidic traits and expressed in Pascals (Pa).
  4. The patch graft according to claim 3, wherein the first inner layer contains a hyaluronane hydrogel layer exhibiting a first viscoelasticity of approximately 50 Pa to approximately 150 Pa.
  5. The patch graft according to claim 3, wherein the backing contains a hyaluronane hydrogel layer exhibiting viscoelasticity from about 600 to about 800 Pa.
  6. The patch graft according to claim 3, wherein the third outer layer comprises a hyaluronane hydrogel layer and the third viscoelastic property is about 200 to about 300 Pa.
  7. The patch graft according to claim 3, wherein the backing comprises silk.
  8. The patch graft according to any one of claims 1 to 7, wherein the mixture of BTSC and ELSMC is depleted of mature mesenchymal cells.
  9. The patch graft according to claim 8, wherein the mixture of BTSC and ELSMC comprises a plurality of organoids formed by the self-assembly of BTSC and ELSMC.
  10. Multiple organoids, a) At least one marker selected from the pluripotent gene group consisting of OCT4, Sox2, Sall4, Nanog, Klf5, Cdx2, and Bmi1, b) At least one marker selected from the endodermal transcription factor group consisting of Sox9, Sox17, Pdx1, HNF4 alpha, HNFB1, and ONECUT2, c) A BTSC that is positive for at least one marker selected from the group of stem cell/precursor-related surface markers consisting of one or more isoforms of EpCAM, NCAM, LGR5, CD44, CXCR4, sodium-iodine cotransporter (NIS), CD49 (integrin A6), CD29 (integrin B1), and integrin B4; BTSCs are negative for markers of mature hepatocytes or pancreatic cells, including P450, aquaporins, enzymes involved in bile production, amylase, and digestive enzymes. The patch graft according to claim 9.

Description

This application, which cross-references related applications, claims priority and benefit from U.S. Application No. 16/422,086, filed on 24 May 2019. Furthermore, this application relates to International Patent Application PCT/US2018/036960, filed on 11 June 2018, which claims priority from U.S. Provisional Patent Application No. 62/518,380, filed on 12 June 2017, and No. 62/664,694, filed on 30 April 2018. The contents of these applications are incorporated herein by reference in their entirety. Sequence Listing This application includes a sequence listing filed electronically in ASCII format, which is incorporated herein by reference in its entirety. This invention generally relates to the field of cell transplantation or tissue engraftment. More specifically, to transplantation from solid organ to solid organ or tissue, particularly to internal organs. The invention relates to compositions and methods that provide strategies for the rapid transplantation, engraftment, and integration of cells into solid organ tissues for the treatment of solid organ diseases or the establishment of disease model systems. Representative examples of this possibility include cell therapy for the treatment of liver or pancreatic diseases. There are unmet needs for transplantation strategies involving solid organ-derived cells, hematopoietic cells, mesenchymal stem cells, or strategies different from those used in skin transplantation. Transplantation of hematopoietic and mesenchymal cells, typically derived from single-cell suspensions, is conventionally performed via vascular channels and relies on the activation of adhesion molecules at the relevant target site due to microenvironmental signaling, a process called "homing." Methods used in skin transplantation employ transplantation techniques using cells and/or tissues applied directly to the target site. Transplantation of cells derived from solid organs other than skin has traditionally been done via vascular routes or by direct injection into the tissue. This approach is ill-advised because adhesion molecules on cells derived from solid organs are constantly activated, leading to rapid (second-by-second) cell aggregation that can generate life-threatening embolisms. Even with embolisms well managed to minimize health risks, cell engraftment efficiency is low, at less than 20% for adult cells and even lower (<5%) for stem cells/progenitors. Most transplanted cells die or are transported to ectopic sites where they can survive for months, generating tissue in inappropriate locations and potentially resulting in clinically adverse effects. The small percentage of cells that engraft at the target site integrate slowly, taking weeks to months to reconstruct critical parts of the tissue. When cells are coated with hyaluronan and delivered via vascular pathways through hyaluronan clearance in the tissue (e.g., liver), improvements are seen in engraftment in the liver and minimizing ectopic cell delivery. However, this improvement is still less efficient than using transplantation strategies and, importantly, still allows for cell delivery to ectopic sites. Grafting solid organs is challenging to design due to concerns regarding the size, shape, and complexity of the organ structure, in addition to dynamic mechanical forces. Therefore, there remains a need to improve methods for cell engraftment into solid organs. This disclosure addresses this need and provides relevant advantages. This specification describes novel patch graft compositions and methods for transplanting cells into tissues and solid organs. In one embodiment, the present disclosure relates to a method for engrafting cells onto a solid organelle of an object that requires them, the method being This involves bringing the patch graft into contact with a solid organ. The patch comprises a mixture of epithelial cells and mesenchymal cells incorporated into a biomaterial having first viscoelastic properties, and the biomaterial facilitates contact between at least a portion of the aforementioned epithelial cells, mesenchymal cells, or both, to promote engraftment between cells of a solid organ. This includes clearly indicating that at least a portion of the aforementioned epithelial cells, mesenchymal cells, or both are engrafted among the cells of a solid organ. In some embodiments, demonstrating that at least a portion of the aforementioned epithelial cells are engrafted among the cells of a solid organ is indicated by measuring the secretion level from the solid organ or the metabolic effect of the solid organ in a biological sample obtained from the subject. In another aspect, the disclosure relates to a method for engrafting cells onto a solid organelle of an object that requires them, the method being This involves bringing the patch graft into contact with a solid organ. The patch comprises a mixture of epithelial cells and mesenchymal cells incorporated into a hydrogel layer having first viscoelastic prop