KR-20260067382-A - Composition and method for the activation and maturation of immune cells

Abstract

Mammals rely on adaptive immunity to protect against infections and tumors, and the thymus plays a crucial role by producing T cells that target infected or cancerous cells. However, the thymus undergoes functional decline with age, resulting in significant impairment of the immune system in the elderly. To investigate ways to reverse this decline, we disclose compositions and methods for activating and differentiating T cell progenitors in extrathymic tissues to stimulate the production of differentiated T cells. For example, LNP-mediated delivery of mRNA encoding DLL-1, IL-7, and FLT3L to the liver is shown to generate transient sites for T cell maturation that enhance T cell-mediated immunity in aged mice without causing autoimmunity. By promoting ectopic T cell development from hematopoietic progenitors and activating dendritic cells, these approaches provide evidence that the manipulation of adaptive immunity in vivo can effectively alleviate immunosenescence.

Inventors

• 장 펑
• 프리드리히 미르코 율리안

Assignees

• 더 브로드 인스티튜트, 인코퍼레이티드
• 매사추세츠 인스티튜트 오브 테크놀로지

Dates

Publication Date

20260512

Application Date

20240802

Priority Date

20230802

Claims (20)

1. (a) One or more polynucleotides encoding one or more activating proteins that induce T cell differentiation of T cell progenitor cells, and (b) A delivery vehicle configured to deliver one or more polynucleotides to a target tissue in which T cell precursors differentiate. A composition comprising
2. A composition according to claim 1, wherein one or more activating proteins are agonists of the Notch signaling pathway or the T cell receptor (TCR) signaling pathway.
3. A composition according to paragraph 2, wherein the agonist of the Notch signaling pathway comprises DLL1, DLL4, Jagged1, Jagged2, or a combination thereof.
4. In paragraph 2, the composition wherein the agonist of the TCR signaling pathway is a major histocompatibility complex (MHC) class I or MHC class II ligand.
5. A composition according to claim 4, wherein the MHC class I ligand comprises HLA-A, HLA-B, HLA-C, or a homolog thereof.
6. A composition according to claim 4, wherein the MHC class II ligand comprises HLA-DP, HLA-DQ, HLA-DR, or a homolog thereof.
7. A composition according to claim 1, further comprising one or more cytokines, or one or more polynucleotides encoding said cytokines.
8. A composition according to claim 7, wherein one or more cytokines comprise IL-7, FLT3L, KIT ligand, IL-2, IL-4, IL-15, CCL25, or a combination thereof.
9. A composition according to claim 7, wherein one or more cytokines induce the expression of transcription factor RUNX3.
10. A composition according to any one of claims 1 to 7, wherein one or more polynucleotides comprise one or more plasmid DNAs encoding one or more activating proteins and/or cytokines; one or more mRNA molecules encoding one or more activating proteins and/or cytokines; or a combination thereof.
11. A composition according to claim 10, wherein one or more mRNAs include one or more modifications that increase mRNA stability, decrease immunogenicity, or improve translation efficiency.
12. A composition according to claim 1, wherein the delivery vehicle comprises lipid nanoparticles, exosomes, microvesicles, viral vectors, engineered retrocomponent vectors, polynucleotide-based nanostructures, or extracellular contractile injection systems.
13. A composition according to claim 12, wherein the viral vector comprises a retrovirus, lentivirus, adenovirus, adeno-associated, or herpes simplex virus vector.
14. A composition according to claim 1, wherein the target tissue is the liver, spleen, small intestine, thymus, or a combination thereof.
15. As a method for inducing T cell precursor differentiation, A step of delivering one or more polynucleotides encoding one or more activating proteins that induce differentiation of T cell precursors to a target tissue where T cell precursor differentiation occurs. A derivation method that includes
16. In paragraph 15, the induction method is such that one or more polynucleotides are formulated to be delivered to endothelial cells of a target tissue so that an activating protein is displayed on the surface of the endothelial cells.
17. In paragraph 15 or 16, the induction method wherein the target tissue is the liver, spleen, small intestine, thymus, or a combination thereof.
18. A method of induction according to any one of claims 15 to 17, wherein one or more activating proteins induce differentiation of T cell precursors into double-positive (DP) T cells.
19. A method of induction according to any one of claims 15 to 18, wherein one or more activating proteins are agonists of the Notch signaling pathway or the T cell receptor (TCR) signaling pathway.
20. In paragraph 19, the induction method comprises an agonist of the Notch signaling pathway including DLL1, DLL4, Jagged1, Jagged2, or a combination thereof.

Description

Composition and method for the activation and maturation of immune cells Cross-reference regarding related applications This application claims the benefit of U.S. Provisional Application No. 63/530,465 filed August 2, 2023, and U.S. Provisional Application No. 63/631,183 filed April 8, 2024. The entire contents of the identified applications are incorporated herein by reference in their entirety. Statement regarding government-funded research This disclosure was made with government support under research grant numbers 5F31CA275339-02 and 1DP2GM146245-01 awarded by the National Institutes of Health. The government retains certain rights to this disclosure. Reference to the electronic sequence list The contents of the electronic sequence list (BROD-5835WP_ST26.xml"; size 1,167,882 bytes, created on August 2, 2024) are incorporated herein by reference in their entirety. Technology field The subject matter disclosed in this specification generally relates to compositions and methods for activating and differentiating T cell precursors in target tissues to stimulate the production of differentiated T cells. The thymus is an essential organ for establishing and maintaining the adaptive immune system, serving as a site for the selection, proliferation, development, and differentiation of T cells. Over time, the thymus undergoes age-related atrophy (i.e., thymic atrophy), resulting in reduced efficient T cell development and decreased production of inexperienced T cells. This can increase susceptibility to opportunistic infections, autoimmunities, and cancer. The inability to establish adaptive immunity can also lead to poor vaccine responses and increased morbidity and mortality. Therefore, there is a need for compositions and methods to stimulate and differentiate T-cell precursors in non-thymic tissues to rejuvenate the adaptive immune system. The citation or identification of any document in this application does not constitute an acknowledgment that such document is available as prior art for the present disclosure. The present disclosure provides a composition and a method for activating and differentiating T cell precursors in a targeted tissue to stimulate the production of differentiated T cells. In one embodiment, the present disclosure provides a composition comprising (a) one or more polynucleotides encoding one or more activating proteins that induce T cell differentiation of T cell precursor cells, and (b) a delivery vehicle configured to deliver one or more polynucleotides to a target tissue in which the T cell precursors differentiate. In one embodiment, one or more activating proteins are agonists of the Notch signaling pathway or the T cell receptor (TCR) signaling pathway. In another embodiment, agonists of the Notch signaling pathway include DLL1, DLL4, Jagged1, Jagged2, or a combination thereof. In another embodiment, agonists of the TCR signaling pathway are major histocompatibility complex (MHC) class I or MHC class II ligands. In another embodiment, MHC class I ligands include HLA-A, HLA-B, HLA-C, or homologs thereof. In another embodiment, MHC class II ligands include HLA-DP, HLA-DQ, HLA-DR, or homologs thereof. In one embodiment, the agonist of the Notch signaling path does not include DDL-1 or DLL4. In one embodiment, the agonist of the Notch signaling path does not include IL-7. In one embodiment, the agonist of the Notch signaling path does not include FLT3L. In one embodiment, the composition further comprises one or more cytokines or a polynucleotide encoding said cytokines. In another embodiment, one or more cytokines comprise IL-7, FLT3L, KIT ligand, IL-2, IL-4, IL-15, CCL25, or a combination thereof. In another embodiment, one or more cytokines induce the expression of transcription factor RUNX3. In one embodiment, one or more polynucleotides of any composition described herein comprise one or more plasmid DNAs encoding one or more activating proteins and/or cytokines; one or more mRNA molecules encoding one or more activating proteins and/or cytokines; or a combination thereof. In another embodiment, one or more mRNAs comprise one or more modifications that increase mRNA stability and/or reduce immunogenicity and/or improve translation efficiency. In one embodiment, the delivery vehicle comprises lipid nanoparticles, exosomes, microvesicles, a viral vector, an engineered retrocomponent vector, a polynucleotide-based nanostructure, or an extracellular contractile injection system. In another embodiment, the viral vector comprises a retrovirus, lentivirus, adenovirus, adeno-associated, or herpes simplex virus vector. In one embodiment, the target tissue is the liver, spleen, small intestine, thymus, or a combination thereof. In one embodiment, the present disclosure provides a method for inducing T cell precursor differentiation, wherein the method comprises the step of delivering one or more polynucleotides encoding one or more activating proteins that induce differentiation of T