Scalable production of standardized extracellular vesicles, extracellular vesicle preparations and uses thereof
Preparations comprising an enriched population of extracellular vesicles (nEVs) having a negatively charged surface, and that are CD81+ and CD9−, are provided. Improved processes and methods for producing an enriched population of nEVs from non-murine cells, especially human origin cells and/or tissues, are disclosed. Therapeutic methods for using the preparations, including for reducing brain inflammation and treatment of various pathologies associated with brain inflammation, such as by intravenous or intranasal administration, are also described. Methods and preparations for reducing brain inflammation associated with traumatic brain injury (TBI) are also disclosed. A method for treating a patient having suffered a mild traumatic injury (mTMI), or concussion, such as a sports-related head injury, is also disclosed. The nEVs are also demonstrated to reduce the expression level of IL-Iβ in brain tissue of an animal having had traumatic brain injury. Methods for improving cognitive function and performance in animals after a traumatic brain injury is also demonstrated using the preparations of nEVs disclosed herein.
1. A method for improving pattern separation function and spatial learning ability in a human subject subsequent to a traumatic brain injury (TBI) comprising:
(a) providing a base-line separation function and spatial learning ability measurement of a traumatic brain injury control subject identified to have had a traumatic brain injury (TBI) to provide base-line traumatic brain injury assessment measurements;
(b) administering a pharmaceutically acceptable preparation comprising an effective amount of an enriched preparation of negatively charged human extracellular vesicles (n-EVs) that do not express surface epitope CD9 (CD9−) to the human subject having had traumatic brain injury to provide a treated subject;
(c) assessing the treated subject for separation function and spatial learning ability to provide post-treatment assessment measurements, and comparing the post-treatment assessment measurements to the base-line traumatic brain injury assessment measurements; and
(d) identifying the treated subject having improved pattern separation function and spatial learning ability where the post-treatment assessment measurements are improved compared to the base-line assessment measurements.
2. The method of claim 1 , wherein the enriched population of negatively charged extracellular vesicles (n-EVs) comprises about 30 g of protein and 15×10 9 negatively charged extracellular vesicles (n-EVs) by weight of the preparation.
3. The method of claim 1 , wherein the negatively charged extracellular vesicles (n-EVs) have a mean size of about 80 nm to about 250 nm.
4. The method of claim 1 , wherein the pharmaceutically acceptable preparation is suitable for intravenous administration or intranasal administration.
5. The method of claim 1 , wherein the pharmaceutically acceptable preparation comprises a therapeutically active agent or a biologically active agent.
6. The method of claim 1 , wherein the negatively charged extracellular vesicles (n-EVs) lack a combination of two or more of the surface epitopes selected from the group consisting of: CD29, CD44, CD49c, CD49f, CD59, CD73, CD90, CD105, and CD166.
7. The method of claim 1 , wherein the negatively charged extracellular vesicles (n-EVs) comprise at least about 50% by volume of the pharmaceutically acceptable preparation.
8. The method of claim 1 , wherein the pharmaceutically acceptable preparation comprises mature TSG-6 mRNA, cytokines, micro-RNA and mitochondria.
9. The method of claim 1 , wherein the pharmaceutically acceptable preparation comprises an injectable preparation.
10. The method of claim 1 , wherein the negatively charged extracellular vesicles (n-EVs) are CD63+ and CD105−.