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US-20260124273-A1 - Composition, Method, And Use of Netrin-1 In Preserving the Bone Marrow Niche to Promote Stem Cell Health

US20260124273A1US 20260124273 A1US20260124273 A1US 20260124273A1US-20260124273-A1

Abstract

Disclosed is a method of using Netrin-1 (NTN1) driven rejuvenation of an aged hematopoietic system based on its shown dependency of reestablishing integrity of aged bone marrow niche and reactivating DNA damage response (DDR). NTN1 is shown as a master regulator of reactivating DNA damage response (DDR) pathways. Every organ/tissue accumulates DNA damage that is an underlying cause of diseases. Reactivating DDR has an extensive effect on treating diseases. NTN1 serves as a therapeutic modality that effectively reverses age-related hematopoietic deficiencies while simultaneously targeting growth and survival of acute myelogenous leukemia (AML). The ability to target and gene correct hematopoietic stem cells (HSC) ex vivo for the treatment of hemoglobinopathies, like thalassemia and sickle cell anemia, is limited due to inability of transducing the correct cell population. NTN1 is shown to maintain and expand bona fide HSCs, opening up the possibility to transduce a true stem cell overcoming previous limitations.

Inventors

  • Jason Mathew BUTLER
  • Pradeep Ramalingam

Assignees

  • HACKENSACK MERIDIAN HEALTH, INC.

Dates

Publication Date
20260507
Application Date
20231003

Claims (20)

  1. 1 . A pharmaceutical composition comprising a therapeutic drug containing an effective dose of netrin-1 (NTN-1) in a pharmaceutically acceptable carrier or vehicle, wherein the netrin- 1 is sufficient to reactivate DNA Damage Response (DDR), resolve DNA damage, and restore functional potential within an aged bone marrow niche and blood stem cells and restore regenerative capacity of an aged hematopoietic system to endure serial chemotherapy regimens.
  2. 2 . The pharmaceutical composition according to claim 1 , wherein the therapeutic drug is given as a systemic infusion administration to a patient.
  3. 3 . The pharmaceutical composition according to claim 1 , wherein treatment of acute myeloid leukemia (AML) with the NTN1 results in a decrease in the expansion of AML cells due to apoptosis.
  4. 4 . The pharmaceutical composition according to claim 3 , wherein the NTN1 antagonizes leukemia by maintaining aged vascular endothelium (VE) and mesenchymal stem cells (MSC) integrity to maintain aged hematopoietic stem cells (HSC), while eradicating AML simultaneously.
  5. 5 . The pharmaceutical composition according to claim 1 , wherein mesenchymal stem cells (MSC) that are supplemented in an ex vivo expansion protocol with the NTN1 maintain and enhance function of expanded young and aged hematopoietic stem cells (HSC) for gene correction of hemaglobinopathies.
  6. 6 . The pharmaceutical composition according to claim 1 , wherein systemic infusion of NTN1 restores aged hematopoietic stem cells (HSC) function by reactivating the DNA Damage Response (DDR), and the NTN1 reactivates DDR to promote rapid hematopoietic recovery, improved self-renewal capacity, increased longevity, and reduces weight loss following single and serial myelosuppressive insults as compared to non-treatment with the therapeutic drug.
  7. 7 . Use of a therapeutic drug which contains an effective dose of netrin-1(NTN-1) for treatment of hemoglobinopathies.
  8. 8 . The use according to claim 7 , wherein the hemoglobinopathies includes thalassemia
  9. 9 . The use according to claim 7 , wherein the NTN1 is able to induce reactivation of DNA damage response (DDR) and rejuvenation of an aged hematopoietic system.
  10. 10 . A method of using a pharmaceutical composition comprising: administrating a therapeutic drug containing an effective dose of netrin-1 (NTN-1) to a patient for providing a NTN1-treatment; and using the NTN1-treatment in preserving a bone marrow (BM) niche for promoting stem cell health as compared to not providing the NTN1-treatment.
  11. 11 . The method of claim 10 , wherein the therapeutic drug is for hematological malignancies and minimizes toxicities associated with strand of care myelosuppressive treatments as compared to conventional treatment.
  12. 12 . The method of claim 10 , further comprises utilizing Recombinant murine NTN1 protein reconstituted in PBS at 100 μg/mL.
  13. 13 . The method of claim 12 , further comprises taking a dosage of 4 μg NTN1; diluting the dosage of 4 μg NTN-1 to a final volume of 100 μL; and injecting the final volume subcutaneously.
  14. 14 . The method of claim 13 , wherein the injecting the final volume negatively affects acute myeloid leukemia (AML) aggressiveness.
  15. 15 . The method of claim 14 , wherein the injecting the final volume rejuvenates stem cells function of supportive niche within bone marrow of mice.
  16. 16 . The method of claim 15 , wherein the injecting the final volume further includes preserving a hematopoietic/vascular function while simultaneously targeting AML cells.
  17. 17 . The method of claim 10 , further comprises performing alkaline comet assays and confirming that the NTN1-treatment resolves DNA damage in aged HSCs to levels observed in young mice.
  18. 18 . The method of claim 17 , further comprises demonstrating that the NTN1-treatment reactivates DNA damage response (DDR) and resolving DNA damage within aged hematopoietic stem cells (HSCs).
  19. 19 . The method of claim 18 , further comprises maintaining HSC homeostatis as compared to not having the NTN1-treatment.
  20. 20 . The method of claim 10 , further comprises utilizing the NTN1-treatment as a therapeutic modality that reverses age-related hematopoietic deficiencies while simultaneously targeting growth and survival of acute myelogenous leukemia (AML) as compared to not utilizing the NTN1-treatment.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS The present application is a U.S. national phase entry under 35 U.S.C. § 371 of International Application No. PCT/US2023/34334, filed, Oct. 3, 2023 which claims the benefit of the filing date of U.S. Provisional Application No. 63/413,033, filed Oct. 4, 2022, entitled Composition, Method, And Use of Netrin-1 In Preserving the Bone Marrow Niche to Promote Stem Cell Health, the disclosure of which is hereby incorporated herein by reference. TECHNICAL FIELD The present disclosure relates to a method, composition, and uses of Netrin-1 and application thereof in rejuvenating aged niche cells and restoring blood stem cell function. The present disclosure also relates to a method composition, and uses of Netrin-1 in maintaining an active DNA damage response (DDR) and method for reversing functional deficits within aged blood stem cells by rejuvenating their supportive niche within bone marrow. BACKGROUND The human body is susceptible to disease every day. Each of the approximately ten trillion cells in the human body receives tens of thousands of DNA lesions per day. These lesions can block genome replication and transcription, and if they are not repaired or are repaired incorrectly, they lead to mutations or wider-scale genome aberrations that threaten cell or organism viability. Some DNA aberrations arise by physiological processes, such as DNA mismatches occasionally introduced during DNA replication and DNA strand breaks caused by abortive topoisomerase I and topoisomerase II activity. In addition, hydrolytic reactions and non-enzymatic methylations generate thousands of DNA-base lesions per cell per day. DNA damage is also produced by reactive-oxygen compounds arising as by-products from oxidative respiration or through redox-cycling events involving environmental toxins and Fenton reactions mediated by heavy metals. Reactive oxygen and nitrogen compounds are also produced by macrophages and neutrophils at sites of inflammation and infections. Such chemicals can attack DNA, leading to adducts that impair base-pairing and/or block DNA replication and transcription, base loss, or DNA single-strand breaks (SSBs). Furthermore, when two SSBs arise in close proximity, or when the DNA-replication apparatus encounters a SSB or certain other lesions, double-strand breaks (DSBs) are formed. While DSBs do not occur as frequently as the other lesions listed above, they are difficult to repair and extremely toxic. Today, probably the most prevalent environmental cancer-causing chemicals are those produced by tobacco products, which trigger various cancers, most notably those of the lung, oral cavity, and adjacent tissues. Cancer-causing DNA-damaging chemicals can also contaminate foods, such as aflatoxins found in contaminated peanuts and heterocyclic amines in over-cooked meats. To combat threats posed by DNA damage, human cells have mechanisms collectively termed the DNA-damage response (DDR)-to detect DNA lesions, signal their presence and promote their repair. Cells defective in these mechanisms generally display heightened sensitivity towards DNA-damaging agents, and many such defects cause human disease. Although responses differ for different classes of DNA lesions, they usually occur by a common general regime. The issue is DDR in cells, especially in aged systems, do not function properly and fail to do the repairs needed thus resulting in a disease state. There is an urgent need to discover how to promote DDR in cells. Netrins are known to regulate neuronal migration, axon guidance and synaptogenesis in the nervous system. Netrins are a class of laminin-like proteins, which were first identified as axonal guidance cues during embryonic development. Netrin-1 protein is membrane associated in the adult spinal cord. Over recent years, it has become apparent that netrin-1 may additionally be involved in the underlying pathology of several multisystem diseases, making it an attractive potential therapeutic target. However, there are conflicting reports as to whether netrin-1 acts in a pro- or anti-angiogenic capacity. For example, in atherosclerosis, opposing effects have been reported on plaque progression, due to the ability of netrin-1 to inhibit both macrophage egress from and monocyte ingress into lesions. Endothelial netrin-1 expression has also been shown to regulate leucocyte migration, and modulation of this could potentially provide benefit in exacerbating or attenuating inflammatory responses. To date, there has been limited research on other proteins in the netrin family, which may also be involved in these processes. Even in the case of netrin-1, further investigation is needed to determine whether it can play a part in the prevention of cardiovascular disease as well as other inflammatory pathologies. The seemingly contradictory actions of netrin-1 in certain physiological pathways, such as angiogenesis, serve to further highlight its intricate and multiple roles