WO2020252287A1 - Enhancement of fibroblast therapeutic activity by rna - Google Patents
Enhancement of fibroblast therapeutic activity by rna Download PDFInfo
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- WO2020252287A1 WO2020252287A1 PCT/US2020/037467 US2020037467W WO2020252287A1 WO 2020252287 A1 WO2020252287 A1 WO 2020252287A1 US 2020037467 W US2020037467 W US 2020037467W WO 2020252287 A1 WO2020252287 A1 WO 2020252287A1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/35—Fat tissue; Adipocytes; Stromal cells; Connective tissues
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- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0656—Adult fibroblasts
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K35/14—Blood; Artificial blood
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- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/33—Fibroblasts
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/36—Skin; Hair; Nails; Sebaceous glands; Cerumen; Epidermis; Epithelial cells; Keratinocytes; Langerhans cells; Ectodermal cells
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/48—Reproductive organs
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- C12N2500/00—Specific components of cell culture medium
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- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/12—Hepatocyte growth factor [HGF]
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- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/90—Polysaccharides
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- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/30—Synthetic polymers
- C12N2533/32—Polylysine, polyornithine
Definitions
- the technical field of the disclosure includes at least the fields of cell biology, molecular biology, cell therapy, and medicine.
- Fibroblasts comprise the main cell type of connective tissue, possessing a spindle- shaped morphology, whose classical function has historically been believed to produce extracellular matrix responsible for maintaining the structural integrity of tissue. Fibroblasts also play an important role in proliferative phases of wound healing, resulting in deposition of extracellular matrix [1, 2]. During wound healing, scar tissue is formed by fibroblast over proliferation. In embryos, and in some types of amphibians, scar-less healing occurs after injury by processes which are currently under intense investigation [3, 4]. With aging, many kinds of tissues and organs undergo fibrosis gradually, such as fibrosis of skin, lung, liver, kidney and heart.
- scar tissue formation is caused by hyperproliferation of fibroblasts, as well as these cells producing abnormally large amounts of extracellular matrix and collagens during proliferation and thereby replacing normal organ structure (parenchyma), leading to functional impairment and scar formation, which may further trigger persistent fibrosis.
- Embodiments of the disclosure include methods and compositions for
- the disclosure there are means of generating fibroblast cells possessing enhanced therapeutic activity after transplantation.
- the disclosure pertains to means of utilizing exposure to an effective amount of RNA as a method of increasing therapeutic activity of fibroblasts.
- administration of double stranded RNA is performed through providing polyinosinic-polycytidylic acid (poly (I:C)) or a functionally active derivative thereof at a concentration sufficient to induce one or more therapeutic properties and/or to augment therapeutic properties onto the fibroblasts.
- enhanced therapeutic activity comprises augmentation of fibroblast migratory activity.
- therapeutic activities are selected from a group comprising of: a) angiogenesis; b) immune modulation; c) differentiation ability; d) production of trophic factors; e) ability to resist apoptosis; f) migratory activity; and g) a combination thereof.
- any composition of the invention may be used in any method of the disclosure, and any method of the disclosure may be used to produce or to utilize any composition of the invention.
- Aspects of an embodiment set forth in the Examples are also embodiments that may be implemented in the context of embodiments discussed elsewhere in a different Example or elsewhere in the application, such as in the Brief Summary, Detailed Description, Claims, and Brief Description of the Drawings.
- FIG. 1 provides an example of a cell migration assay.
- Triangles refer tp Poly EC, “X” refers to CpB, and squares are scrambled RNA; and
- FIG. 2 shows HGF production from fibroblasts following exposure to Poly (I:C). Bars from left to right are Control, low molecular weight Poly (I:C), and high molecular weight Poly (I:C).
- the terms“or” and“and/or” are utilized to describe multiple components in combination or exclusive of one another.
- “x, y, and/or z” can refer to“x” alone,“y” alone,“z” alone,“x, y, and z,”“(x and y) or z,”“x or (y and z),” or“x or y or z.” It is specifically contemplated that x, y, or z may be specifically excluded from an
- exogenous refers to RNA that originated from outside the fibroblast cells.
- the subject can be any organism or animal subject that is an object of a method or material, including mammals, e.g., humans, laboratory animals (e.g., primates, rats, mice, rabbits), livestock (e.g., cows, sheep, goats, pigs, turkeys, and chickens), household pets (e.g., dogs, cats, and rodents), horses, and transgenic non-human animals.
- the subject can be a patient, e.g., have or be suspected of having a disease (that may be referred to as a medical condition), such as one or more infectious diseases, one or more genetic disorders, one or more cancers, one or more chronic medical conditions, one or more injuries, or any combination thereof.
- a disease that may be referred to as a medical condition
- the disease may or may not be pathogenic.
- the subject may being undergoing or having undergone antibiotic treatment.
- the subject may be asymptomatic.
- the subject may be healthy individuals.
- “individual”, as used herein, may or may not be housed in a medical facility and may be treated as an outpatient of a medical facility.
- the individual may be receiving one or more medical compositions via the internet.
- An individual may comprise any age and any gender of a human or non-human animal and therefore includes both adult and juveniles ( i.e ., children) and infants and includes in utero individuals. It is not intended that the term connote a need for medical treatment, therefore, an individual may voluntarily or involuntarily be part of experimentation whether clinical or in support of basic science studies.
- Embodiments of the disclosure include methods of enhancing one or more therapeutic activities of a fibroblast population comprising the steps of: a) optionally selecting a fibroblast population; and b) treating a fibroblast population with a concentration of RNA sufficient to enhance one or more therapeutic properties of the fibroblasts.
- the RNA may or may not be double stranded RNA, such as double stranded RNA being polyinosine-polycytidylic acid (Poly (I:C)).
- the double stranded RNA may be polyinosine-polycytidylic acid stabilized with Polylysine and Carboxymethylcellulose (Poly ICLC).
- the fibroblasts may be derived from a source selected from the group consisting of a) adipose tissue; b) dermal tissue; c) placental tissue; d) hair follicles; e) keloid tissue; f) bone marrow; g) peripheral blood; h) umbilical cord; i) foreskin; and j) a combination thereof.
- compositions of cells may comprise any of the fibroblasts encompassed herein.
- the preparations may or may not also comprise RNA, such as Poly (I:C).
- any method may or may not include the step of inducing activation of toll like receptor 3, for example through contact with a ligand capable of inducing an interferon response in the fibroblast cells.
- the method may further comprise the step of delivering a therapeutically effective amount of the cells to an individual at risk of having a medical condition or that has a medical condition for which the cells would be therapeutic, such as remove or reduce the severity of at least one symptom.
- the disclosure discloses the previously unknown and paradoxical properties of the use of RNA molecules to enhance therapeutic activity of fibroblast cells.
- RNA molecules are utilized in a sequence non-specific, and/or sequence semi specific manner in order to activate molecular pathways inside fibroblasts capable of inducing production of one or more interferons.
- the disclosure provides that the production of one or more interferons is associated with enhanced ability of the fibroblasts to migrate towards an area of injury, for example, towards a SDF-1 gradient that could be associated with an injury.
- poly (I:C) may be formulated with one or several bioadhesive polymers that can prolong the residence time in tissue culture, in order to maintain fibroblast activation.
- micro particles are particles with an average particle size between 0.1 mhi and 100 mhi.
- the carrier polymer is starch, such as obtained from maize, potato or cassava.
- nanoparticles may be utilized for delivery of Poly (I:C) to fibroblasts in vitro.
- the term Growth Medium generally refers to a medium sufficient for the culturing of umbilicus-derived cells.
- one particular medium for the culturing of the cells of the disclosure herein comprises Dulbecco's Modified Essential Media (also abbreviated DMEM herein).
- DMEM-low glucose also DMEM-LG herein
- the DMEM-low glucose may be supplemented with 15% (v/v) fetal bovine serum (e.g ., defined fetal bovine serum, Hyclone, Logan Utah), antibiotic s/antimycotic s (such as penicillin (100 Units/milliliter), streptomycin (100 milligrams/milliliter), and amphotericin B (0.25 micrograms/milliliter), (Invitrogen, Carlsbad, Calif.)), and 0.001% (v/v) 2-mercaptoethanol (Sigma, St. Louis Mo.). In some cases different growth media are used, or different
- supplementations are provided, and these are normally indicated in the text as supplementations to Growth Medium.
- the fibroblast cells may be cultured in standard growth conditions.
- standard growth conditions refers to culturing of cells at 37° C., in a standard atmosphere comprising 5% CO2. Relative humidity is maintained at about 100%. While foregoing the conditions are useful for culturing, it is to be understood that such conditions are capable of being varied by the skilled artisan who will appreciate the options available in the art for culturing cells, for example, varying the temperature, CO2, relative humidity, oxygen, growth medium, and the like.
- fibroblasts treated with RNA are utilized to treat one or more inflammatory conditions.
- the term“inflammatory conditions” includes, for example: (1) tissue damage due to ischemia-reperfusion following acute myocardial infarction, aneurysm, stroke, hemorrhagic shock, crush injury, multiple organ failure, hypovolemic shock intestinal ischemia, spinal cord injury, and traumatic brain injury; (2) inflammatory disorders, e.g., bums, endotoxemia and septic shock, adult respiratory distress syndrome, cardiopulmonary bypass, hemodialysis; anaphylactic shock, severe asthma, angioedema, Crohn's disease, sickle cell anemia, poststreptococcal glomerulonephritis, membranous nephritis, and pancreatitis; (3) transplant rejection, e.g., hyperacute xenograft rejection; (4) pregnancy related diseases such as recurrent fetal loss and pre-eclampsi
- Complement-mediated inflammation associated with autoimmune disorders including, but not limited to, myasthenia gravis, Alzheimer's disease, multiple sclerosis, rheumatoid arthritis, systemic lupus erythematosus, insulin-dependent diabetes mellitus, acute disseminated encephalomyelitis, Addison's disease, antiphospholipid antibody syndrome, autoimmune hepatitis, Crohn's disease, Goodpasture's syndrome, Graves' disease, Guillain-Barre syndrome, Hashimoto's disease, idiopathic thrombocytopenic purpura, pemphigus, Sjogren's syndrome, and Takayasu's arteritis, may also be treated with the methods described herein.
- fibroblasts treated with RNA are used to treat one or more neurodegenerative conditions.
- A“neurodegenerative condition” encompasses acute and chronic conditions, disorders or diseases of the central or peripheral nervous system.
- a neurodegenerative condition may be age-related, or it may result from injury or trauma, or it may be related to a specific disease or disorder.
- Acute neurodegenerative conditions include, but are not limited to, conditions associated with neuronal cell death or compromise including cerebrovascular insufficiency, e.g. due to stroke, focal or diffuse brain trauma, diffuse brain damage, spinal cord injury or peripheral nerve trauma, e.g., resulting from physical or chemical burns, deep cuts or limb severance.
- Examples of acute neurodegenerative disorders are: cerebral ischemia or infarction including embolic occlusion and thrombotic occlusion, reperfusion following acute ischemia, perinatal hypoxic-ischemic injury, cardiac arrest, as well as intracranial hemorrhage of any type (such as epidural, subdural, subarachnoid and intracerebral), and intracranial and intravertebral lesions (such as contusion, penetration, shear, compression and laceration), as well as whiplash and shaken infant syndrome.
- cerebral ischemia or infarction including embolic occlusion and thrombotic occlusion, reperfusion following acute ischemia, perinatal hypoxic-ischemic injury, cardiac arrest, as well as intracranial hemorrhage of any type (such as epidural, subdural, subarachnoid and intracerebral), and intracranial and intravertebral lesions (such as contusion, penetration, shear, compression and laceration), as well
- Chronic neurodegenerative conditions include, but are not limited to, Alzheimer's disease, Pick's disease, diffuse Lewy body disease, progressive supranuclear palsy (Steel-Richardson syndrome), multisystem degeneration (Shy-Drager syndrome), chronic epileptic conditions associated with neurodegeneration, motor neuron diseases including amyotrophic lateral sclerosis, degenerative ataxias, cortical basal degeneration, ALS-Parkinson's-Dementia complex of Guam, subacute sclerosing panencephalitis, Huntington's disease, Parkinson's disease, synucleinopathies
- the fibroblasts for use in any method of the current disclosure may be of any mammalian origin, e.g., human, rat, primate, porcine and the like.
- the fibroblasts are derived from human umbilicus.
- Umbilicus-derived cells are capable of self-renewal and expansion in culture, and have the potential to differentiate into cells of other phenotypes.
- Methods of deriving cord tissue fibroblast cells from human umbilical tissue are contemplated.
- the cells are capable of self-renewal and expansion in culture, and have the potential to differentiate into cells of other phenotypes.
- the method comprises one or more steps of (a) obtaining human umbilical tissue; (b) removing substantially all of blood to yield a substantially blood-free umbilical tissue, (c) dissociating the tissue by mechanical or enzymatic treatment, or both, (d) re-suspending the tissue in a culture medium, and (e) providing growth conditions which allow for the growth of a human umbilicus -derived cell capable of self-renewal and expansion in culture and having the potential to differentiate into cells of other phenotypes.
- Tissue can be obtained from any completed pregnancy, term or less than term, whether delivered vaginally, or through other routes, for example surgical Cesarean section. Obtaining tissue from tissue banks is also considered within the scope of the present invention.
- the tissue is rendered substantially free of blood by any means known in the art.
- the blood can be physically removed by washing, rinsing, and diluting and the like, before or after bulk blood removal for example by suctioning or draining.
- Other means of obtaining a tissue substantially free of blood cells might include enzymatic or chemical treatment.
- Dissociation of the umbilical tissues can be accomplished by any of the various techniques known in the art, including by mechanical disruption, for example, tissue can be aseptically cut with scissors, or a scalpel, or such tissue can be otherwise minced, blended, ground, or homogenized in any manner that is compatible with recovering intact or viable cells from human tissue.
- the isolation procedure also utilizes an enzymatic digestion process.
- Many enzymes are known in the art to be useful for the isolation of individual cells from complex tissue matrices to facilitate growth in culture.
- a broad range of digestive enzymes for use in cell isolation from tissue is available to the skilled artisan. Ranging from weakly digestive (e.g ., deoxyribonucleases and the neutral protease, dispase) to strongly digestive (e.g., papain and trypsin), such enzymes are available commercially.
- weakly digestive e.g ., deoxyribonucleases and the neutral protease, dispase
- strongly digestive enzymes e.g., papain and trypsin
- a nonexhaustive list of enzymes compatable herewith includes mucolytic enzyme activities, metalloproteases, neutral proteases, serine proteases (such as trypsin, chymotrypsin, or elastase), and
- deoxyribonucleases Presently considered are enzyme activities selected from metalloproteases, neutral proteases and mucolytic activities.
- collagenases are known to be useful for isolating various cells from tissues.
- Deoxyribonucleases can digest single- stranded DNA and can minimize cell-clumping during isolation.
- Enzymes can be used alone or in combination.
- Serine protease are preferably used in a sequence following the use of other enzymes as they may degrade the other enzymes being used. The temperature and time of contact with serine proteases must be monitored. Serine proteases may be inhibited with alpha 2 microglobulin in serum and therefore the medium used for digestion may be serum-free.
- EDTA and DNase are commonly used and may improve yields or efficiencies.
- Particular methods involve enzymatic treatment with for example collagenase and dispase, or collagenase, dispase, and hyaluronidase, and such methods are provided wherein in certain preferred embodiments, a mixture of collagenase and the neutral protease dispase are used in the dissociating step.
- Particular methods include those methods that employ digestion in the presence of at least one collagenase from Clostridium histolyticum, and either of the protease activities, dispase and thermolysin. Still more preferred are methods employing digestion with both collagenase and dispase enzyme activities.
- enzyme treatments are known in the art for isolating cells from various tissue sources.
- the LIB ERASE BLENDZYME (Roche) series of enzyme combinations of collagenase and neutral protease are very useful and may be used in the instant methods.
- Other sources of enzymes are known, and the skilled artisan may also obtain such enzymes directly from their natural sources.
- the skilled artisan is also well-equipped to assess new, or additional enzymes or enzyme combinations for their utility in isolating the cells of the invention.
- Certain enzyme treatments may be 0.5, 1, 1.5, or 2 hours long or longer.
- the tissue is incubated at 37° C. during the enzyme treatment of the dissociation step. Diluting the digest may also improve yields of cells as cells may be trapped within a viscous digest.
- the cells can be re-suspended after the tissue is dissociated into any culture medium as discussed herein above.
- Cells may be re-suspended following a centrifugation step to separate out the cells from tissue or other debris. Resuspension may involve mechanical methods of re- suspending, or simply the addition of culture medium to the cells.
- Providing the growth conditions allows for a wide range of options as to culture medium, supplements, atmospheric conditions, and relative humidity for the cells.
- a particular temperature is 37° C., however the temperature may range from about 35° C. to 39° C.
- methods of deriving umbilical cells capable of expansion in the absence of particular growth factors are similar to the method above, however they require that the particular growth factors (for which the cells have no requirement) be absent in the culture medium in which the cells are ultimately re-suspended and grown in. In this sense, the method is selective for those cells capable of division in the absence of the particular growth factors.
- Particular cells in some embodiments are capable of growth and expansion in chemically-defined growth media with no serum added. In such cases, the cells may require certain growth factors, which can be added to the medium to support and sustain the cells.
- the number of units to be allocated depends on the number of nucleated cells in each unit and the medical condition to be treated. In some embodiments, the amount of fibroblasts, or the number of units, available for allocation to an individual depends on the amount of donations made.
- the present disclosure provides a method of fibroblast banking, the method comprising periodically collecting a plurality of donations from an individual throughout the individual's life.
- the method comprises collecting fibroblasts from more than one source.
- the method comprises collecting fibroblasts of more than one type.
- lentiviral based transfection of genes into fibroblasts include transfection of SDF-1 to promote stem cell homing, particularly hematopoietic stem cells [12], GDNF to treat Parkinson’s in an animal model [13], HGF to accelerate remyelination in a brain injury model [14], akt to protect against pathological cardiac remodeling and cardiomyocyte death [15], TRAIL to induce apoptosis of tumor cells [16-19], PGE-1 synthase for cardioprotection [20], NUR77 to enhance migration [21], BDNF to reduce ocular nerve damage in response to hypertension [22], HIF-1 alpha to stimulate osteogenesis [23], dominant negative CCL2 to reduce lung fibrosis [24], interferon beta to reduce tumor progression [25], HLA-G to enhance immune suppressive activity [26], hTERT to induce differentiation along the hepatocyte lineage [27], cytosine deaminase [28], OCT-4 to reduce senescence [29, 30],
- Cells were assessed for chemotaxis to the indicated chemokine (SDF-1) under normoxic conditions for 2 h (FIG. 1). Migrated cells were collected from the lower migration chamber compartments and counted. Cells were seeded at 2.5 x 10 6 /mL in the upper chamber of a Transwell system (3 mm pore size; Coming Costar, 3415). 10% FBS RPMI 1640 medium alone or supplemented with recombinant human CXCL12 (100 ng/mL) (Peprotech, 300-28A), or CCL19 (0.3 pg/mL), or CCL21 (0.6 pg/mL) was added to the lower compartment. Cells were allowed to migrate for 2 h at 37°C under normoxic condition. Cells migrated in the lower chamber were collected and counted.
- SDF-1 indicated chemokine
- Fibroblasts were cultured as in Example 1 and treated with control, low, or high molecular weight Poly (I:C) from InvivoGen® (San Diego, CA). Cells were cultured for 48 hours and HGF concentration was assessed using ELISA. Substantial stimulation of HGF production was noted with both high and low molecular weight Poly (I:C) (FIG. 2). HGF is one example of a cytokine that mediates stem cell therapeutic effects.
- fibroblasts produce enhanced production of one or more cytokines, such as HGF, following exposure to an effective amount of Poly (I:C), when compared to fibroblasts that were not exposed to the cytokine(s), such as HGF.
- cytokines such as HGF
- HGF HGF-like growth factor
- Therapeutic properties of HGF include: stimulation of liver regeneration, stimulation of renal tubular epithelial cell proliferation, enhancement of recovery of renal function after injury, stimulation of keratinocyte growth, stimulation of angiogenesis, inhibition of cancer cell proliferation, stimulation of hematopoiesis, enhances B cell activity, stimulation of bronchial epithelial cell growth, stimulation of type 2 alveolar epithelial cells, inhibitory of epithelial cell apoptosis, stimulation of lung healing, reduction of pulmonary fibrosis, enhancement of pancreatic regeneration, promotes survival of neurons, promotes growth of axons, activation of muscle satellite cells, accelerates reconstitution of intestinal epithelial cells, accelerate post cardiac infarct recovery, suppresses cardiomyopathy, inhibits autoimmune myocarditis, reduces endothelial cell injury, reduces graft versus host disease, reduction of stroke size and acceleration of recovery, suppression of neuronal death, increases brain hypoperfusion, inhibits progression of neurodegenerative diseases,
- Glavaski-Joksimovic, A., et al. Glial cell line-derived neurotrophic factor-secreting genetically modified human bone marrow-derived mesenchymal stem cells promote recovery in a rat model of Parkinson's disease. J Neurosci Res, 2010. 88(12): p. 2669- 81.
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JP2021573328A JP2022536664A (en) | 2019-06-12 | 2020-06-12 | Enhancement of fibroblast therapeutic activity by RNA |
AU2020292370A AU2020292370A1 (en) | 2019-06-12 | 2020-06-12 | Enhancement of fibroblast therapeutic activity by RNA |
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WO2021207192A1 (en) * | 2020-04-08 | 2021-10-14 | Figene, Llc | Methods and compositions for allergy and asthma treatment using fibroblasts |
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US20070237747A1 (en) * | 2003-05-28 | 2007-10-11 | Peter Angel | Method for Increasing the Yield of Proliferating Primary Keratinocytes |
US20150225699A1 (en) * | 2014-02-10 | 2015-08-13 | The Board Of Trustees Of The Leland Stanford Junior University | ACTIVATION OF INNATE IMMUNITY FOR ENHANCED NUCLEAR REPROGRAMMING OF SOMATIC CELLS WITH mRNA |
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EP2665708A4 (en) * | 2011-01-20 | 2014-07-09 | Univ Colorado Regents | Modulators of tlr3/dsrna complex and uses thereof |
WO2016019472A1 (en) * | 2014-08-08 | 2016-02-11 | Quest Pharmatech Inc. | Tumor antigen specific antibodies and tlr3 stimulation to enhance the performance of checkpoint interference therapy of cancer |
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US20070237747A1 (en) * | 2003-05-28 | 2007-10-11 | Peter Angel | Method for Increasing the Yield of Proliferating Primary Keratinocytes |
US20150225699A1 (en) * | 2014-02-10 | 2015-08-13 | The Board Of Trustees Of The Leland Stanford Junior University | ACTIVATION OF INNATE IMMUNITY FOR ENHANCED NUCLEAR REPROGRAMMING OF SOMATIC CELLS WITH mRNA |
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WO2021207192A1 (en) * | 2020-04-08 | 2021-10-14 | Figene, Llc | Methods and compositions for allergy and asthma treatment using fibroblasts |
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AU2020292370A1 (en) | 2022-02-03 |
US20220235326A1 (en) | 2022-07-28 |
EP3982984A4 (en) | 2023-07-12 |
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JP2022536664A (en) | 2022-08-18 |
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