The existing CAR-engineered T cell-based (CAR-T) therapy represents one of the most successful immunotherapy approaches developed in recent years. Most CAR-T cell therapy has been used clinically to treat hematological malignancies by targeting the B cell-specific antigen, CD19. However, this approach is not without limitations due to toxicities such as by neurotoxicity or cytokine release syndrome. Additionally, CAR-T cells function only as autologous cells due to graft-versus-host disease that would develop if cells were obtained from another person. Therefore, CAR-T cells must be produced on a patient-specific basis. NK cells, on the other hand, function as allogenic cytotoxic effector cells that do not have to be utilized on a patient-specific basis and are proven to be less toxic since they do not cause cytokine release syndrome, neurotoxicity, or graft-versus-host disease. For these reasons, CAR-engineered NK (CAR-NK) cells have increasingly attracted interest as an alternative CAR-cell therapy. However, there exist other unmet challenges. Targeting CAR-based therapies against solid tumors has been challenging due to the lack of truly tumor-specific antigens as most targets are shared by non-malignant cells and can cause toxicity due to “on-target, off-tumor” effects.” A fine-tunable CAR therapy is useful to better identify and target tumors while limiting this toxicity.

During a normal blood loss injury, the body forms a clot to stop bleeding. Forming a blood clot requires certain blood cells, platelets, and their interactions with protein clotting or coagulation factors. Bleeding disorders comprise a group of conditions resulting when blood cannot clot properly due to defects in the platelets or clotting factors, and may range from mild to life threatening. Hemophilia is a more well-known inherited X-linked bleeding disorder, predominantly affecting males, and current incidence is estimated at about 1/5000 live male births. Patients with hemophilia A lack clotting factor VIII (FVIII), and patients with Hemophilia B lack clotting factor IX (FIX). Patients manifest predominantly with severe spontaneous joint and muscle bleeds and often die from intracranial bleeding. Treatment and prophylaxis of bleeds consist of frequent infusions with FVIII or FIX clotting factor concentrates. Approximately 30% of hemophilia patients develop inhibitors (neutralizing antibodies) to FVIII or FIX. These patients are treated with FVIIa-based bypassing agents, but treatment remains suboptimal. Bypassing agents do not restore the normal pathways of hemostasis in hemophilia, but rather enable clotting by boosting thrombin generation and avoid the inhibitor’s blocking effects by circumventing a requirement for FVIII or FIX. However, at least 1/3 of patients do not respond adequately to FVIIa-based products, leaving these patients unprotected against bleeding, with a high burden of joint disease and at high risk of fatal bleeding.In some cases patients are administered anticoagulant medications that may be associated with an increased risk of bleeding.  

Mounting evidence suggests that dormant leukemia stem cells (LSC) evade therapies that target cycling cells. On the basis of the observation that sonic hedgehog (Shh) signaling pathways modulate cell cycle regulation in normal hematopoiesis, inventors hypothesized that forcing dormant refractory cells back into the cell cycle might also make them vulnerable to drugs that target actively dividing cells.

This invention provides microRNA therapeutics that augment blood vessel growth, which may have application for indications where it is desired to reduce or stimulate angiogenesis. Reducing or inhibiting angiogenesis may be useful for indications such as degenerative eye diseases and cancer. Stimulating blood vessel growth may be useful for treating indications such as cardiovascular, thrombotic or ischemic diseases. Cells lining blood vessels are usually among the least proliferative cell types, but this desired quiescence may be interrupted in response to growth factors during pathological neovascularization manifested in disease states such as macular degeneration and cancer. MicroRNAs are known to be key regulators of angiogenesis and specific miRNAs have been found to be effective toward these indications.

Septic shock occurs from an overwhelming bacterial infection and is characterized by severe hypotension with low blood flow. It is the 13th leading cause of death in the United States with a mortality rate of 30%-50% due to the lack of effective treatments. In addition to the devastating effects of this syndrome on individuals, it incurs billions of dollars annually in healthcare costs.