Treatment of cachexia using fibroblast cells and products thereof

Abstract

To provide methods and compositions related to treatment of cachexia or inflammation related thereto.SOLUTION: In specific embodiments, cachexia or inflammation related thereto is to treated or prevented by the administration of fibroblasts. In specific embodiments, the fibroblasts are amniotic-derived fibroblasts.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] This application is a continuation of U.S. Provisional Patent Application No. 62 / 755,542, filed November 4, 2018. No. 6,396,433, filed on Oct. 23, 2003, which is incorporated by reference in its entirety.

[0002] (Technical field) This disclosure relates to at least the fields of cell biology, molecular biology, cell therapy, and medicine. be. [Background technology]

[0003] Cachexia is a muscle wasting disorder that is particularly prevalent in patients with cancer or chronic infections such as HIV. It is a wear-and-tear-related condition. Inflammatory mediators such as TNF-α are responsible for cachexia. Studies have shown that interventions to inhibit inflammation are relatively ineffective in clinical settings. One potential reason for this discrepancy is the multifactorial nature of cancer-associated inflammation.

[0004] The process of neoplastic transformation is associated with the uncontrolled proliferation of cells, which leads to de novo Necrosis of vo tissue occurs, resulting in macrophage infiltration, complement activation, and neovascularization. The development of this neoplasm occurs at a level that interferes with normal physiological processes. In these conditions, a systemic increase in inflammatory markers has been observed, which may be due to a systemic effect on the patient's overall condition. This results in excessive chronic inflammation.

[0005] The majority of cancer patients die from their disease due to muscle wasting, metabolic disorders, and multiple organ failure. Currently available interventions focus on reducing tumor burden in cancer patients; The present invention relates to a method for reducing pathology associated with the effects of a tumor mass on the general physiology of a patient. There is a need for interventions that improve physiological, immunological and metabolic outcomes in cancer patients. The present invention provides a method for using fibroblasts to restore metabolic homeostasis, This allows the tumor to initiate, maintain and execute activities that result in tumor stabilization and possibly regression. This allows for the possibility of innate and adaptive host responses to Summary of the Invention

[0006] An embodiment of the present disclosure includes administering to an individual an effective amount of immunomodulatory fibroblasts. The present invention also includes a method for ameliorating cachexia or inflammation associated therewith in an individual. also induces the production of immunomodulatory activity, thereby producing immunomodulatory fibroblasts. The method may include culturing a population of fibroblasts under suitable conditions for the treatment of cachexia or Related inflammation includes chronic infections, tuberculosis, HIV / AIDS, cancer, abdominal disease, and chronic obstructive pulmonary disease. , multiple sclerosis, rheumatoid arthritis, congestive heart failure, familial amyloid polyneuropathy , mercury poisoning (acrodynia), Crohn's disease, untreated / severe type 1 diabetes, anorexia nervosa chronic inflammation, such as one selected from the group consisting of rheumatoid arthritis, hormonal deficiency, and combinations thereof; In some cases, a chronic inflammatory state may be initiated by inflammation that is detected in the bloodstream. The individual is associated with an elevation of one or more detectable markers, such as one or more markers. Known to have or suspected to have a chronic inflammatory condition Examples of detectable markers include C-reactive protein, Interleukin-1, interleukin-6, TNF-α, and combinations of these Examples of the compound include those selected from the group consisting of:

[0007] In certain embodiments of the present disclosure, the fibroblasts utilized in the methods are derived from amniotic fluid. Examples of fibroblasts derived from human, primate, canine, feline, caprine, elephant, bovine, equine, porcine The animal may be selected from the group consisting of mice, rabbits, and combinations thereof. In certain embodiments, the fibroblasts are SSEA3, SSEA4, Tra-1-60, T ra-1-81, Tra-2-54, Oct-4, CD13, CD44, CD49b, C D105, aminopeptidase N, and combinations thereof. In certain instances, the fibroblasts express detectable levels of one or more markers. do not express extractable levels of SSEA1.

[0008] When an individual is provided with fibroblasts, they may be administered epidurally, intracerebroventricularly, intraventricularly, subcutaneously, sublingually, or intravenously. , outside the cheek-amniotic membrane, nose, intra-arterial, intra-cardiac, intracavernosal, intradermal, intra-lesional, intra-muscular, intra-ocular, intra-bone, Intraperitoneal, intrathecal, intrauterine, intravaginal, intravenous, intravesical, intravitreal, subcutaneous, transdermal, perivascular, transcutaneous cachexia, by one or more routes selected from the group consisting of: mucosal, and combinations thereof or to an individual suffering from inflammation associated therewith.

[0009] The present disclosure relates to fibroblasts having regenerative properties, tissue associated cells having regenerative properties, and Cell-derived products with regenerative properties for the treatment of cachexia or inflammation associated therewith In one embodiment, the present invention relates to a method for treating cancer-associated cachexia or HIV-associated cachexia (e.g., ) is a regenerative medicine that has been modified to allow for administration while retaining the properties associated with cell regeneration. Treatment is by administration of cells or tissues bearing the cells. Administration may optionally be intravenous. The mode of administration may be subcutaneous, intramuscular or intravenous.

[0010] In an embodiment of the present disclosure, a method for improving cachexia or inflammation associated therewith in an individual is provided. There is a method for inducing the production of an immunomodulatory activity, comprising the steps of: a. Culturing a population of fibroblasts under appropriate conditions, thereby producing immunomodulatory fibroblasts. and b. administering to the individual an effective amount of said immunomodulatory fibroblasts. In some cases, cachexia or the associated inflammation is a cause of chronic infection, tuberculosis, HIV / AIDS, cancer, and abdominal cavity. Disease, chronic obstructive pulmonary disease, multiple sclerosis, rheumatoid arthritis, congestive heart failure, familial amyloidosis Idiopolyneuropathy, mercury poisoning (acromialgia), Crohn's disease, untreated / severe type 1 diabetes , anorexia nervosa, hormone deficiency, and combinations thereof. In certain embodiments, the chronic inflammatory condition is initiated by a chronic inflammatory condition such as C-reaction. responsive proteins, interleukin-1, interleukin-6, TNF-α, and and combinations thereof. In certain embodiments, fibroblasts are derived from amniotic fluid (e.g., , humans, primates, dogs, cats, goats, elephants, cows, horses, pigs, mice, rabbits, and other animals. The amniotic fluid is derived from a species selected from the group consisting of the following combinations: fibroblasts The cells were SSEA3, SSEA4, Tra-1-60, Tra-1-81, and Tra-2- 54, Oct-4, CD13, CD44, CD49b, CD105, aminopeptidase N, and a detectable level of one or more markers selected from the group consisting of N, N, and combinations thereof. In certain embodiments, the ibulocochlear cells may express detectable levels of SSE. Does not express A1.

[0011] Fibroblasts can be injected epidurally, intraventricularly, intracerebroventricularly, subcutaneously, sublingually, extrabuccally-amniotically, nasally, intraarterially, or cardiacly. Intravisceral, intracavernous, intradermal, intralesional, intramuscular, intraocular, intraosseous, intraperitoneal, intrathecal, intrauterine, intravaginal, Intravenous, intravesical, intravitreal, subcutaneous, transdermal, perivascular, transmucosal, and combinations thereof. The method according to any one of claims 1 to 10, wherein the composition is administered by one or more routes selected from the group consisting of: The method described.

[0012] The above has been presented to illustrate the features and technical advantages of the present disclosure in order that the following detailed description may be better understood. Having outlined rather broadly, additional features and advantages form the subject of the claims of this specification. The disclosed concepts and specific embodiments are intended to accomplish the same objectives of the present design. Those skilled in the art will appreciate that the present invention may be readily utilized as a basis for modifying or designing other structures to achieve the same object. It should be understood by those skilled in the art that such equivalent constructions are within the scope of the appended claims. It should be understood by those skilled in the art that the present invention is not intended to depart from the spirit and scope of the present invention. Novel features believed to be characteristic of the designs disclosed herein include further objectives and The following general description, together with its advantages, both as to organization and method of operation, may be considered in conjunction with the accompanying drawings, in which: The above and other aspects of the present invention will be better understood from the following description. However, the figures are presented for purposes of illustration and explanation only. It should be expressly understood that these are provided for purposes of illustration only and are not intended as a definition of the limits of the present disclosure. It is. [Brief description of the drawings]

[0013] For a more complete understanding of the present disclosure, reference is now made to the following descriptions taken in conjunction with the accompanying drawings, in which:

[0014] [Figure 1]Treatment of cachexia with skin foreskin fibroblasts. The weight of certain mice exposed to the different conditions was measured. From left to right for each of the four bar groups: 1) control mice subcutaneously injected with PBS in the right flank of the axilla (Ctrl group); 2) cachectic mice subcutaneously injected with 1x106 LLC cells in the right flank of the axilla (CA group); 3) cachectic mice treated with 100,000 foreskin fibroblasts (cachexia+fibroblasts(1)); and 4) 200,000 foreskin fibroblasts (cachexia+fibroblasts(2)). Detailed Description of the Invention

[0015] I. Definition

[0016] In accordance with long-standing patent law convention, the words "a" and "an" refer to the invention inclusive of the claims. As used herein, "one or more" means one or more of the following: Consisting of, or consisting essentially of, one or more elements, method steps, and / or methods of the disclosure. Any of the methods or compositions described herein may be used in combination with any of the methods or compositions disclosed herein. may be implemented with respect to any other method or composition disclosed herein and still fall within the spirit and scope of the disclosure. It is contemplated that similar or similar results can be obtained without departing from the scope of the claims. Use of the word "or" in a clause refers to alternatives only, unless expressly stated. "And / or" is used to mean that the alternatives are mutually exclusive, but the disclosure supports definitions that refer only to alternatives and "and / or".

[0017] Throughout this specification, unless the context otherwise requires, the words "include" and "include" are used interchangeably. "Contains," "includes," and "comprises" mean the inclusion of a specified set of steps or elements, but not others. It will be understood that the exclusion of any step, element or group of elements is not intended. does not mean excluding elements. "Consists of" means "consists of" the following: Thus, "consisting of" does not indicate that the listed elements are required or mandatory. "Consists of" indicates that other elements may be present. "Consists essentially of" means that any element may be present. "includes" means that the listed elements are required or essential, but other elements are optional, It may or may not be present depending on whether it affects the activity or action of the described element. This also shows that it is good.

[0018] Throughout this specification, the terms "one embodiment," "embodiment," "particular embodiment," "related embodiment," "relevant embodiment," "relevant" and "relevant" are used interchangeably. "particular embodiment", "additional embodiment", or "further embodiment" References to specific features, configurations, or combinations thereof described in connection with the embodiments are not intended to be construed as limiting the present invention. or features are included in at least one embodiment of the present invention. The appearances of the above phrases in various places throughout this specification do not necessarily all refer to the same embodiment. Moreover, any particular feature, structure or characteristic may be incorporated into one or more embodiments. may be combined in any suitable manner.

[0019] As used herein, the terms "or" and / or "and" and "and / or" Terms are used to describe multiple elements in combination with one another or exclusively, e.g. For example, "x, y, and / or z" means "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" In particular, x, y, or z may be specifically excluded from the embodiment. It is specifically intended that

[0020] Throughout this application, the term "about" refers to a value that is relative to the device or method used to determine the value. In order to show that the standard deviation of the error for Used according to its simple and ordinary meaning.

[0021] "Subject" and "subject" and "individual" refer to any human or animal, including primates, mammals, and vertebrates. In certain embodiments, the subject is a human, a dog, a cat, a horse, or a non-human. For example, cattle.

[0022] "Treating" or treatment of a disease or condition means alleviating at least one sign or symptom of the disease. The process may include administering one or more drugs or treatments (including cells) to the patient to The desired effect of treatment is to slow the rate of disease progression. to reduce, ameliorate or alleviate the condition, or delay the onset of at least one symptom Relief includes the reduction of symptoms before the onset of a disease or condition, and the reduction of symptoms before the onset of a disease or condition. The term "treatment" can refer to a condition that may occur before, during, or after the onset of the condition, or both. "Preventing" or "preventing" a disease or undesirable condition can include "preventing" or "preventing" a disease or undesirable condition. Further, "treat" or "treating" refers to the complete alleviation of one or more signs or symptoms. A protocol that does not require surgery, does not require healing, and specifically has only a minor effect on the patient. Including calls.

[0023] As used herein, the term "prevent" and its variants such as "prevent" and "prevent" are used interchangeably. Synonyms refer to approaches to prevent, inhibit, or reduce the likelihood of occurrence or recurrence of a disease or condition. For example, delaying the onset or recurrence of cancer, a disease or condition, or preventing or treating a disease or condition. or, as used herein, "prevent" and "delay the onset or recurrence of symptoms of Synonyms refer to the intensity, effect, symptoms, and / or severity of a disease or condition prior to the onset or recurrence of the disease or condition. This also includes reducing the load.

[0024] As used herein, the term "pharmacologically acceptable" refers to an animal or does not produce adverse, allergic, or other untoward reactions when administered to humans; It refers to the fruiting body and composition.

[0025] As used herein, the term "pharmaceutical acceptable carrier" refers to any suitable carrier, including water, ethanol, polyvinyl chloride, ethyl acetate ... Polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol) oils, coatings, isotonic and absorption retarding agents, etc.), and suitable mixtures thereof. Any suitable antibacterial agent, including but not limited to, antibacterial agents, liposomes, commercially available detergents, etc. Any and all solvents or dispersion media are included. Supplementary biologically active ingredients may also be used in such compositions. The signal processing unit 100 may be incorporated into a carrier.

[0026] Cachexia is the involuntary weight loss associated with cancer, infection, and other catabolic states. Cachexia is a condition that cannot be cured by nutritional supplementation. Various chronic diseases such as liver disease, chronic obstructive pulmonary disease (COPD), HIV, and rheumatoid arthritis In patients with idiopathic rheumatoid arthritis, it is often associated with a terminal course. Other symptoms include anorexia, malabsorption, nausea, asthenia, neuroendocrine changes, immune system dysfunction, energy Additional symptoms include disturbances in glucose metabolism. Several methods for treating cachexia have been proposed. Although there are suggestions that it may be possible to treat the disease, the focus has been on developing treatments for the underlying disease. There is no direct treatment for the fluid.

[0027] Recently, the multifactorial nature of cachexia has been highlighted, particularly the role of inflammatory mediators in the initiation and perpetuation of this condition. There has been some success in better understanding the role of the Bacterial lipopolysaccharide (LPS), or endotoxin, is a pathogenic factor in bacterial infections, sepsis, and critical illness. Originating from early studies implicating the pathophysiological expression of monocytes in macrophage / monocyte lineage-derived Cytokines can mimic many of the effects of endotoxin and contribute to the clinical phenotype of cachexia. It was observed that ribosomal protein is a central mediator of oxidative stress.

[0028] The present disclosure relates to the use of fibroblasts as a means of reducing cachexia in individuals in need of treatment. More particularly, the present disclosure relates to the use of More particularly, the present disclosure relates to the use of fibroblasts in treating cachexia in patients in need of treatment. The present invention provides a method for increasing the activity of specific cells to prevent or treat the following conditions: The individual may be an aging patient, the individual may be a cancer patient, the individual may be a patient suffering from a virus or other The individual may be a patient infected with an infectious disease and / or a medical condition having cachexia as a symptom. The individual may be a patient having a chronic disorder or disease. The individual may be of any age, sex, or race. It is possible.

[0029] In certain embodiments, the present disclosure provides a method for inhibiting cachexia or reducing at least one symptom of cachexia. Regenerative cells, in one embodiment, fibroblasts, for reducing the severity and / or delaying the onset of cachexia. In certain embodiments, administering an effective amount of fibroblasts to an individual provides Fibroblasts can suppress tumor-derived molecules that can induce muscle atrophy and immune suppression.

[0030] Embodiments of the present disclosure provide a method for ameliorating cachexia or inflammation associated therewith in an individual. A method for inducing the production of immunomodulatory activity, thereby producing immunomodulatory fibroblasts. and administering to the individual an effective amount of an immunization agent. The method includes administering regulatory fibroblasts.

[0031] In certain embodiments, an effective amount of fibroblasts, including immunomodulatory fibroblasts, is provided to an individual. Increasing weight in an individual with cachexia or at risk of cachexia, comprising There are methods of preventing weight loss in an individual. In certain embodiments, the fibroblasts are SEA3, SSEA4, Tra-1-60, Tra-1-81, Tra-2-54, Oc t-4, CD13, CD44, CD49b, CD105, aminopeptidase N, and The present invention relates to a method for producing a detectable level of one or more markers selected from the group consisting of the above combinations. Manifest.

[0032] The present disclosure provides a method for inhibiting inflammation, thereby suppressing the host body's ability to induce muscle wasting. The present invention includes any type of fibroblast that is capable of secreting progeny, such as foreskin-derived fibroblasts. The cells are immortal in culture, maintain euploidy for over a year in culture, and resemble human embryonic stem cells. They share one or more markers with stem cells and / or are associated with one of the three germ layers of the developing embryo: endoderm, mesoderm, It can differentiate into all germ layers and ectoderm.

[0033] In one embodiment, the fibroblasts are derived from amniotic fluid, where the cells are from the first trimester of a human pregnancy. Amniotic fluid is extracted from amniotic membranes harvested during the second trimester. Amniotic fluid contains multiple morphologically distinct cell types. It is known that a large proportion of cells are prone to senescence and are lost from culture. No. 7,569,385, the entirety of which is incorporated herein by reference. Using fibronectin-coated plates and culture conditions as described in Cells are grown from amniotic fluid collections from 8 weeks gestation. The cells of the present disclosure are of fetal origin and are normal. The karyotype may be diploid. The growth of amniotic fluid-derived fibroblasts is multipotent, since several major cell types are derived from them. The term "pluripotent" means that amniotic fluid regenerative cells can give rise to several major cell types. This refers to the ability to differentiate, and amniotic fluid-derived fibroblasts proliferate under certain conditions and become "pluripotent." The term "pluripotent stem cells" refers to stem cells that, when cultured under conditions, give rise to specific cell types. Describes stem cells that can differentiate into any type of somatic cell. Amniotic fluid regenerative cells are derived from humans. However, amniotic fluid regenerative cells can be isolated in a similar manner from other species. Examples of species that can be used to derive live cells include mammals, humans, primates, dogs, cats, etc. , goats, elephants, endangered species, cows, horses, pigs, mice, rabbits, etc. The present invention is not limited to these.

[0034] Amniotic fluid-derived fibroblasts are characterized by their specific cell surface proteins or at least In specific cases, they can be recognized by the presence of specific cellular proteins. Certain cell types have specific cell surface proteins. These surface proteins are responsible for the These can be used as "markers" to determine or confirm the cell type. Surface markers can be visualized using antibody-based techniques or other detection methods. Surface markers of isolated amniotic fluid-derived fibroblasts were identified using monoclonal antibodies and FACS analysis. The amniotic fluid samples were extracted from independently collected samples and analyzed for a series of cell surface and other markers. These cells can be characterized by the following cell surface markers: SSEA3, SSEA4, Tra-1-60, Tra-1-81, Tra-2-54, or Fibroblasts express the cell surface marker SSEA1, whereas mesenchymal cells express the cell surface marker SSEA1. Fibroblasts can be distinguished from mesenchymal cells in that they do not express stem cell transcription factor O Fibroblasts may, in certain embodiments, express the following cell markers: at least one, or at least two, or at least three, or at least four, or Can be recognized by at least five, or at least six, or all of the entities: Some For: SSEA3, SSEA4, Tra-1-60, Tra-1-81, Tra-2- 54 and / or Oct-4.

[0035] In certain embodiments, the fibroblasts of the present disclosure in culture express little or no SSEA-1 marker. Amniotic fluid-derived fibroblasts express little or no expression of the embryonic stem cell marker SSEA3, In addition to SSEA4, Tra1-60, Tra1-81, Tra2-54, and Oct-4, Cell surface proteins commonly found on human mesenchymal stem cells, but not on human embryonic stem cells The antigen was also expressed at high levels (MF Pittinger et al., Science 284: 143-147, 1999;S Gronthos et al., J. Cell Physiol. 189:54-63, 2001). This set of markers includes CD13 (99.6%) and Aminopyrimidine dehydrogenase N, CD44 (99.7%) hyaluronan-binding receptor, CD49b (9 9.8%) collagen / laminin-binding integrin α2, and CD105 (97%) endonucleases Embryonic stem cell marker and hMSC marker on fibroblast cultures The presence of both indicates that amniotic fluid-derived fibroblasts grown and expanded as described herein are human stem cells. 13A-C represent a novel class of cells.

[0036] In some embodiments of the present disclosure, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% fibroblasts In some embodiments, at least about 90% of the cells in the culture express CD13. , 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 1 In some embodiments of the present disclosure, at least 100% of the cells in culture express CD44. At least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, In certain embodiments of the present disclosure, the culture At least about 90%, 91%, 92%, 93%, 94%, 95%, 96% of the cells in the culture , 97%, 98%, 99%, or 100% express CD105. Morphology: At least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 5%, 96%, 97%, 98%, 99%, or 100% express SSEA3. In certain embodiments, at least about 90%, 91%, 92%, 93% of the cells in the culture , 94%, 95%, 96%, 97%, 98%, 99%, or 100% express SSEA4 In some embodiments of the present disclosure, at least about 90%, 91% of the cells in the culture , 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% In certain embodiments of the present disclosure, at least one of the cells in culture expresses Tra-1-60. Also about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% %, or 100% of the cultures express Tra-1-81. At least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 9 7%, 98%, 99%, or 100% of the cells express Tra-2-54. In embodiments, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% express Oct-4. In certain embodiments shown, at least about 90%, 91%, 92%, 93%, or 100% of the cells in the culture are %, 94%, 95%, 96%, 97%, 98%, 99%, or 100% aminopeptides It expresses enzyme N.

[0037] A "nutrient medium" is a medium for culturing cells that contains nutrients that promote growth; Nutrient media may be utilized in embodiments of the present disclosure. The appropriate combination of either acid, antibiotics, serum or serum replacement, and exogenously added factors is used. It may be included in combination.

[0038] The present disclosure relates to stem cell differentiation techniques that may be useful for differentiating amniotic fluid regenerative cells into fibroblasts. General methods for this can be found in popular textbooks, e.g., Teratocarcinoma and Embryonic Stem Cells: A Practical Approach. Roach (E. J. Robertson, ed., IRL Press Ltd. 1987 );Technical Handbook (PM Wasserman et al. eds., Academic Press ess 1993); In vitro embryonic stem cell differentiation (MVWiles,Meth.22 5:900, 1993); Characteristics and uses of embryonic stem cells: applications to human biology and gene therapy. Perspectives for use (P.D. Rathjen et al., Reprod. Dev. 10:31, 1998 ); and stem cell biology (LM Reid, Curr. Opinion cell Biol. 2:121, 1990), each of which is incorporated herein by reference in its entirety. Can be enjoyed.

[0039] In one embodiment of the present disclosure, fibroblasts are used to treat aging-related cachexia. In another embodiment, the cachexia-associated factor is used to treat cachexia. The dose of fibroblasts provided to a patient of interest is then used to guide the dosage of fibroblasts provided to a patient of interest. In the present study, the fibroblasts used include amniotic fluid-derived fibroblasts, and in some cases In another embodiment, the amnion-derived fibroblasts are administered intravenously. The amniotic membrane can be used as a source of fibroblasts by direct administration. In the present study, fibroblasts are administered intravenously at a concentration of 10 to 200 million cells.

[0040] Administration of cachexia inhibiting cells to a mammalian patient can be by any method, including but not limited to, intravenous, intradermal, or intravenous administration. Transdermal, subcutaneous, intramuscular, inhalation (e.g., via aerosol), oral (e.g., sublingual), topical localized (i.e., both on the skin and on mucosal surfaces, including respiratory tract surfaces), intrathecal, intra-articular, multiple, intracerebral , intra-arterial, intraperitoneal, oral, intralymphatic, intranasal, rectal or vaginal administration, by local catheter This can be by any route, including perfusion, or direct intralesional injection.

[0041] In certain embodiments, the compositions of the present disclosure are administered for a defined period of time (e.g., 0.5 to 2 hours). The compositions of the present disclosure are administered by intravenous pump or intravenous infusion given over a period of time (10 min). The product can be delivered by peristaltic means or in the form of a depot, but any given The preferred route in any given case will depend on the species, age, sex, and overall survival of the subject, as is well known in the art. Factors such as the physical condition, the nature and severity of the condition being treated, and / or the particular composition being administered. In certain embodiments, the route of administration is once a week. once or twice, via bolus or continuous infusion over a period of time.

[0042] In certain embodiments, the route of administration is one or more sites (e.g., thigh, lower back, buttocks, arm). In one embodiment, the compositions of the present disclosure are administered by subcutaneous injection, optionally once or twice weekly. The compositions and / or methods may be administered on an outpatient basis. Those skilled in the art will appreciate that dosages may be adjusted to suit the age, sex, and / or condition of the subject. The clinical course of the disease is based on many factors, including the type, species, and condition (e.g., cachexia activity or associated inflammation). and the desired degree of cachexia or inflammation associated therewith to be treated. It will be appreciated that the effective amount of the composition of the present disclosure can be determined by one of ordinary skill in the art. Doses derived from in vitro or animal model (e.g., cotton rat or monkey) test systems It can be extrapolated from the response curve.

[0043] Examples of dosing regimens that can be used in the methods of the present disclosure include daily, three times a week (intermittent These include, but are not limited to, once a day, once a week, or every 14 days. In this case, the dosing regimen may include, but is not limited to, monthly dosing or dosing every 6-8 weeks. Those skilled in the art will appreciate that dosages are generally higher and / or more effective in initial treatment compared to maintenance therapy. They will likely understand that more frequent dosing is required for treatment.

[0044] In some embodiments, the individual at the onset of a medical condition associated with cachexia is The methods of the present disclosure are substantially similar to one or more treatments for a medical condition. In some cases, symptoms of cachexia may or may not be provided to the individual at the same time. If detectable, the individual is subjected to the methods of the present disclosure. The therapeutic agent may be provided to an individual before, during, and / or after any treatment for a cardiac condition.

[0045] An embodiment of the present disclosure includes a method for determining whether an individual is suffering from weight loss or cachexia. (1) a fibroblast population; (2) a tissue containing a fibroblast population; and / or (3) a fibroblast cell population. and administering to an individual an effective amount of a product produced by the fibroblast population, and / or tissue comprising the fibroblast population. and administering to a patient suffering from cachexia or inflammation associated therewith. Cachexia may or may not be associated with muscle wasting. It may or may not be associated with other conditions such as cancer, inflammatory conditions, chronic infections, tuberculosis, or HIV. In at least some cases, individuals may have chronic inflammation-related The patient is shown to have an elevation of one or more markers. Examples of markers associated with chronic inflammation include, for example, Peripheral blood C-reactive protein, interleukin-1, interleukin-6, and / or Increased levels of TNF-α were observed in fibroblasts compared with age-matched controls. Although they may be derived from any source, in certain embodiments, they are derived from foreskin. In some cases, fibroblasts can adhere to plastic. Blast cells appear to stimulate upregulation of growth factor production, which stimulates anti-cachexia activity. The amniotic tissue may be administered to an individual in the form of minced amniotic tissue, which may or may not be treated with a steroid. In certain cases, such treatment may involve hypoxic, acidic, hypotonic or other conditions. The fibroblasts are administered in the form of a single cell suspension. This may or may not be the case.

[0046] In certain disclosed methods, fibroblasts are used to treat muscle wasting (with or without cachexia), metabolic It is necessary to prevent, delay the onset of, or reduce the severity of, organ dysfunction, and multiple organ failure. The use of serotonin in individuals with muscle wasting, cachexia, metabolic disorders, and / or multiple organ failure is If there is a delay in the disease, the delay may be, for example, on the order of weeks, years, or months.

[0047] The amount of any type of cell for administration to an individual will depend on the severity and / or severity of the cachexia being treated. The amount of the cell injected may depend on the type of cell being injected for treatment. The cells may be administered in a pharma- ceutically acceptable carrier. For example, the composition may be prepared for administration in an isotonic solution of sterile saline. In one embodiment, the pharma- ceutically acceptable carrier is a FAS ligand, an IL-2R, an IL-2, IL-4, IL-8, IL-10, IL-20, IL-35, HLA-G, PD-L1, I-309, IDO, iNOS, CD200, galectin 3, sCR1, arginase, PGE-2, aspirin, lovastatin, pravastatin, simvastatin, pitavastatin Chin, rapamycin, IVIG, naltrexone, TGF-β, VEGF, PDGF-4 , anti-CD45RB antibody, hydroxychloroquine, leflunofin, dicyanogold, sulfa Salazine, methotrexate, glucocorticoids, etanercept, adalimumab, Nakinra, cetanercept-tin, golimumab, infliximab, rituximab, sik rosporin, IFN-γ, everolimus, rapamycin, VEGF-1, FGF-2, amyloidosis one or more additional agents, such as endodopoietin, HIF-1-α, or a combination thereof may include.

[0048] In one embodiment of the present disclosure, fibroblasts are administered by injection (intramuscular injection), including to hypoxic areas. The compound may be administered to a subject by any suitable route, including by injection. These include intravenous, subcutaneous, intrathecal, oral, rectal, intrathecal, intrareticulum, intraventricular, intrahepatic, and intrarenal. do.

[0049] In some embodiments, the fibroblasts are derived from skin, heart, blood vessels, bone marrow, skeletal muscle, liver, pancreas, etc. The tissue may be derived from tissues including brain, adipose tissue, foreskin, placenta, and / or umbilical cord. In morphology, the fibroblasts may be placental, fetal, neonatal, or adult, or a mixture thereof.

[0050] The number of administrations of cells to an individual will depend, at least in part, on the factors described herein. and can be optimized using routine methods in the art. In other embodiments, multiple administrations of cells are required. However, this system is sensitive to the specific needs of individuals, cell loss or other factors that may change over time and circumstances. depends on variables such as the rate of loss of cell activity as a result of the activity of individual cells. Thus, each individual can be monitored for appropriate dosage, and such monitoring of the individual It is expected that such practices are routine in the art.

[0051] In one embodiment of the present disclosure, cells (e.g., fibroblasts) are cultured to enhance cell viability and To preserve the proliferation and proliferation potential of the cells, the cells are cultured ex vivo using means known in the art. In certain embodiments of fibroblasts, the fibroblasts are directed against the recipient immune system. to achieve one or more desired effects on the cells, such as reducing the visibility of There may be modifications of known culture techniques. In one embodiment, cells (e.g., fibroblasts) are cultured in a manner similar to that described above. ) are cultured under conditions lacking one or more xenogeneic components (e.g., fetal bovine serum). In embodiments, the present disclosure provides a method for the preparation of a cellular antibody, such as a fibroblast, for example, to reduce the immunogenicity of a cell. Additional characterization of fetal bovine serum was performed using human platelet-rich plasma, platelet lysate, umbilical cord serum, and autologous This includes replacing serum and / or a defined cytokine mixture. EXAMPLES

[0052] The following examples are included to demonstrate certain non-limiting aspects of the disclosure. The techniques disclosed in the examples are believed to work well in the practice of the disclosed subject matter. It should be understood by those skilled in the art that it represents technology discovered by the inventors. However, those of skill in the art will appreciate that in light of this disclosure, many of the specific embodiments disclosed may be Modifications may be made, similar or implied, without departing from the spirit and scope of the disclosed subject matter. It should be understood that similar results can still be obtained. Example 1 Treatment of cachexia using forehead fibroblasts

[0053] Lewis Lung Carcinoma (LLC) cells were cultured from the American Type Culture Collection (ATCC). and supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Waltham, MA, USA) and 1% penicillin / streptomycin (Boster Biolabs, Inc., Waltham, MA, USA). The medium was incubated at 5% CO in a 37-well plate with Dulbecco's modified Eagle's medium (G-Technology, Wuhan, China). 2 Prior to tumor inoculation, LLC cells were incubated at 1000 rpm for 5 min at 4 °C. The cells were centrifuged and resuspended in phosphate buffered saline (PBS).

[0054] Specific pathogen-free C57BL / 6 mice, aged 5–6 weeks, were cultured in a specific pathogen-free medium. The mice were kept in a solid diet in a warm environment with a normal 12-hour light-dark cycle and stable temperature and humidity conditions. C57BL / 6 mice were randomly divided into three groups (7 mice per group). The mice were divided into two groups: 1) control mice that were subcutaneously injected with PBS in the right flank of the axilla (Ctrl group); 2) axillary 1 × 10 in the right flank of the fossa 6 Cachectic mice (CA group) subcutaneously injected with LLC cells of 1 Cachexia treated with 0.000 foreskin fibroblasts (ATCC) (cachexia + fibroblasts (1)) and 4) 200,000 foreskin fibroblasts (cachectic + fibroblasts (2)). The body weight and cumulative food intake of mice (all 7 mice in each group) were recorded 3 days after tumor implantation. We monitored the following:

[0055] Although the present disclosure and its advantages have been described in detail, the scope of the present invention is defined by the appended claims. Various changes, substitutions, and alterations may be made herein without departing from the spirit and scope of the present design. It is understood that the scope of the present application is limited to the The present disclosure is not intended to be limited to particular embodiments of the process, machine, manufacture, composition of matter, means, methods and steps. As will be readily understood from this disclosure, those skilled in the art will appreciate that the present disclosure does not intend to perform substantially the same function or achieve substantially the same result as the corresponding embodiment(s) disclosed herein. Any now existing or later developed process, machine, manufacture, composition of matter, means, Methods or processes may be utilized in accordance with the present disclosure. The scope of the present invention includes within its scope such processes, machines, manufacture, compositions of matter, means, methods, or processes. It is intended to include processes.

Claims

[Claim 1] 1. A method for ameliorating cachexia or inflammation associated therewith in an individual, comprising the steps of: a. optionally culturing a population of fibroblasts under appropriate conditions to induce the production of an immunomodulatory activity, thereby producing immunomodulatory fibroblasts; and b. Administering to the individual an effective amount of immunomodulatory fibroblasts.

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