Method for reversing hair loss and the loss of cuticle growth and density by administering effective courses of granulocyte-colony stimulating factor

Abstract

The present invention provides a method for re-growing hair in patients with androgenic alopecia by administering effective consecutive courses of Granulocyte-Colony Stimulating Factor or derivatives. After the state of re-growth is obtained, the hair growth is maintained by administering periodic courses of Granulocyte-Colony Stimulating Factor or derivatives. The invention further provides a method for increasing cuticle growth and density using a similar administration of effective consecutive courses of Granulocyte-Colony Stimulating Factor or derivatives. The increased cuticle growth and density is maintained by administering periodic courses of Granulocyte-Colony Stimulating Factor or derivatives.

Claims

1. A method for re-growing hair in a patient with hair loss by administering effective consecutive courses of Granulocyte-Colony Stimulating Factor, wherein said Granulocyte-Colony Stimulating Factor is selected from a group consisting of Granulocyte-Colony Stimulating Factor, its derivative, functionally similar biological or chemical compound or combination thereof.

2. The method of claim 1, wherein the effective consecutive courses of Granulocyte-Colony Stimulating Factor to re-growth hair are two or more consecutive courses repeated about every 3 to 8 days, wherein said consecutive course administers a daily dose of Granulocyte-Colony Stimulating Factor for about 3 to 6 days.

3. The method of claim 2, wherein said daily dose of Granulocyte-Colony Stimulating Factor administers a Granulocyte-Colony Stimulating Factor dose of about 0.1 micrograms to 100 micrograms per kg body weight per day of Granulocyte-Colony Stimulating Factor on each day.

4. The method of claim 2, wherein said Granulocyte-Colony Stimulating Factor daily dose is administered selected from a group consisting of subcutaneous injection, transdermal patch, intravenously, and orally.

5. The method of claim 1, further comprising providing intravenously adult stem cells to increase hair growth and density upon administration of the effective consecutive courses of Granulocyte-Colony Stimulating Factor to re-growth hair.

6. The method of claim 1, further comprising maintaining hair growth by a periodic effective course of Granulocyte-Colony Stimulating Factor after said consecutive courses are completed, wherein said treatment maintains the hair growth.

7. The method of claim 6, wherein said periodic effective course of Granulocyte-Colony Stimulating Factor to said maintain hair growth is a course of Granulocyte-Colony Stimulating Factor administered daily for about 3 to 6 days and said course is repeated every 2 to 18 weeks, wherein daily dose of Granulocyte-Colony Stimulating Factor is a dose of about 0.1 micrograms to 100 micrograms per kg body weight per day of Granulocyte-Colony Stimulating Factor on each day.

8. The method of claim 7, wherein said administered is selected from a group consisting of subcutaneous injection, transdermal patch, intravenously, and orally.

9. A method for increasing cuticle growth and density by administering effective consecutive courses of Granulocyte-Colony Stimulating Factor, wherein said Granulocyte-Colony Stimulating Factor is selected from a group consisting of Granulocyte-Colony Stimulating Factor, its derivative, functionally similar biological or chemical compound or combination thereof.

10. The method of claim 9, wherein said effective consecutive courses of Granulocyte-Colony Stimulating Factor for increasing cuticle growth and density are two or more consecutive courses repeated about every 3 to 8 days, wherein said consecutive course administers a daily dose of Granulocyte-Colony Stimulating Factor for about 3 to 6 days.

11. The method of claim 10, wherein said daily dose of Granulocyte-Colony Stimulating Factor administers a Granulocyte-Colony Stimulating Factor dose of about 0.1 micrograms to 100 micrograms per kg body weight per day of Granulocyte-Colony Stimulating Factor on each day.

12. The method of claim 9, wherein said cuticle is a fingernail or a toe nail.

13. The method of claim 10, wherein said Granulocyte-Colony Stimulating Factor daily dose is administered selected from a group consisting of subcutaneous injection, transdermal patch, intravenously, and orally.

14. The method of claim 9, further comprising providing intravenously adult stem cells to increase hair growth and density upon administration of the effective consecutive courses of Granulocyte-Colony Stimulating Factor for increasing cuticle growth and density.

15. The method of claim 1, further comprising maintaining cuticle growth and density by a periodic effective course of Granulocyte-Colony Stimulating Factor after said consecutive courses are completed, wherein said treatment maintains the cuticle growth and density.

16. The method of claim 15, wherein said periodic effective course of Granulocyte-Colony Stimulating Factor to maintain cuticle growth and density is a course of Granulocyte-Colony Stimulating Factor administered daily for about 3 to 6 days and said course is repeated every 2 to 18 weeks, wherein daily dose of Granulocyte-Colony Stimulating Factor is a dose of about 0.1 micrograms to 100 micrograms per kg body weight per day of Granulocyte-Colony Stimulating Factor on each day.

17. The method of claim 15, wherein said administered is selected from a group consisting of subcutaneous injection, transdermal patch, intravenously, and orally.

18. A method to maintain hair growth by administering periodic effective courses of Granulocyte-Colony Stimulating Factor, wherein said Granulocyte-Colony Stimulating Factor is selected from a group consisting of Granulocyte-Colony Stimulating Factor, its derivative, functionally similar biological or chemical compound or combination thereof.

19. A method for maintain cuticle growth and density by administering periodic effective courses of Granulocyte-Colony Stimulating Factor, wherein said Granulocyte-Colony Stimulating Factor is selected from a group consisting of Granulocyte-Colony Stimulating Factor, its derivative, functionally similar biological or chemical compound or combination thereof.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0031] FIG. 1 demonstrates the long term reversal of Parkinson's Disease symptoms using the United Parkinson's Disease Scale to evaluate the condition of the patient administered with Granulocyte-Colony Stimulating Factor over two and one-half years.

[0032] FIG. 2 demonstrates the reduction of blood glucose levels below the preferred normal limit after Granulocyte-Colony Stimulating Factor was administered to Type 2 diabetic patient.

[0033] FIG. 3 shows the average time course for blood glucose levels of six treatments with GCSF.

[0034] FIG. 4 shows a GCSF treated identical twin and untreated identical twin with different hair losses.

DETAILED DESCRIPTION OF INVENTION

[0035] The present invention described herein was aimed at performing treatments using Granulocyte-Colony Stimulating Factors administered to patients with neurodegenerative diseases, more preferably Parkinson's disease, to determine whether Granulocyte-Colony Stimulating Factors were able to maintain the state of the diseases, and preferably reverse the symptoms, either partially or completely. Unexpectedly, it was found that the treatment of the neurodegenerative disease, Parkinson's Disease, with Granulocyte-Colony Stimulating Factors reversed symptoms of both the central nervous system (CNS) and peripheral nervous system (PNS). In addition, it was found that Granulocyte-Colony Stimulating Factors delivered in an effective dose was able to reduce blood sugar levels of patients with adult onset Type 2 diabetes for a significant period of time.

[0036] The nucleic acid sequence and encoded amino acid sequence of Granulocyte-Colony Stimulating Factor (GCSF), also referred to as human pluripotent granulocyte colony-stimulating factor, as well as chemically synthesized polypeptides sharing its biochemical and immunological properties has been previously disclosed (U.S. Pat. Nos. 6,379,661; 6,004,548; 6,830,705; 5,676,941, 6,027,720; 5,994,518; 5,795,968; 5,214,132; 5,218,092; 6,261,550; 4,810,643; 4,810,321). Other examples of Granulocyte-Colony Stimulating Factor include analogs which retained their three-dimensional structures and hybrid molecules maintaining their biological and structural integrity were described by Osslund (U.S. Pat. No. 6,261,550). Examples of functional GCSF variants include any proteins, peptides or fragments thereof that are at least 70%, preferably 80% and most preferably at least 90% identity to full-length human GCSF amino acid sequence or its nucleotide sequence. Modifications of GCSF to improve functionality or resident serum clearance include but are not limited to polyethyleneglycol and polyethyleneglycol derivatives thereof, glycosylated forms (Lenogastrim) (WO 00/44785), norleucine analogs (U.S. Pat. No. 5,599,690), addition of amino acids at either terminus to improve folding, stability or targeting, and fusion proteins, such as GCSF and albumin fusion protein (Albugranin) (U.S. Pat. No. 6,261,250). An increase in biological or functional activity over the native peptide may reduce the amount of dose and I or the time period required for treatment. Any chemical or biological entity that functions similar to GCSF can also be employed. GCSF, or the drug name Filgrastim, is currently being sold as Neupogen and its polyethylene glycol modified or pegulated form, with the drug name Pegfilgrastim, sold as Neulasta.

[0037] Examples of closely related functional forms include Granulocyte-Macrophage Colony Stimulating Factor (GMCSF) whose coding DNA sequence and protein including amino acid sequence are known as well as various methods employed to produce recombinant proteins (U.S. Pat. No, 5,641,663). Examples of functional GMCSF variants include any proteins, peptides or fragments thereof that are at least 70%, preferably 80% and most preferably at least 90% identity to full-length human GMCSF amino acid sequence or its nucleotide sequence. Modifications of GCSF to improve functionality or resident serum clearance include but are not limited to polyethyleneglycol and polyethyleneglycol derivatives thereof, glycosylated forms, norleucine analogs, addition of amino acids at either terminus to improve folding, stability or targeting, and fusion proteins, such as GCSF and albumin fusion protein (Albugranin). An increase in biological or functional activity over the native peptide may reduce the amount of dose and/or the time period required for treatment. Any chemical or biological entity that functions similar to GMCSF can also be employed. Examples of GMCSF, or the drug name Sargramostim, which are currently being sold, include Leukine or Leucomax and Leucotropin.

[0038] In one embodiment oral dosages, and methods thereof, of Granulocyte-Colony Stimulating Factor have been described by Nomura and Kazutoshi (U.S. Pat. No. 5,597,562) that allow for dosage reductions, facilitate dose control, and increase the practical usefulness of the bioactive proteins. In addition, Brimelow and Nanette (U.S. Pat. No. 6,497,689) have described preferred pH ranges comprising sulfate ions for stabilizations.

[0039] Granulocyte-Colony Stimulating Factors has been suggested to act as a neuroprotective agent in vitro and may be used for the potential treatment of diseases that result from oxidative stress or apoptosis such as in cerebral ischemia and traumatic brain injury (U.S. Pub. No. 2004/0141946). The work was focused on an in vitro model using STAT proteins and GCSF receptor or rat model for cerebral ischemia. It was suggested that GCSF may be used to treat broadly ischemic or hypoxic related diseases as well as neurological, psychiatric and neurodegenerative diseases as neuroprotective agent acting to slow, interrupt, arrest or stop the progression of the disease. However the work fails to provide insights on how treatment would work in humans as no human study was performed.

[0040] Type 2 diabetes is an example of a non-neurological disorder that appears to be due to the lack of insulin sensitivity of the target cells or insufficient levels of insulin in response to blood glucose. Although it is unknown what causes Type 2 diabetes, it is clear that the disease is usually first diagnosed as an adult and is usually progressive (in terms of the need for therapy to control blood sugar). Type 2 diabetics will have had a relatively long period of time with normal blood sugar before the fasting blood sugar levels begin to rise and the disease can be diagnosed. Other non-neurological disorders include osteoarthritis and benign prostate hypertrophy.

[0041] In view of ongoing research work on stem cells, the use of Granulocyte-Colony Stimulating Factors are likely to enhance or be required in the treatment of neurodegenerative diseases by stem cell therapy. For example, a method for the differentiation of stein cells in culture using Granulocyte-Colony Stimulating Factor and other factors, including lipopolysaccharides, to obtain immune system suppressor cells and immune systems stimulator cells was described by Ogle et al (U.S. Pat. No. 6,165,785). Using a similar approach, a method is useful in differentiating stern cells that are destined to become replacements for damaged cells in neurodegenerative disease and diabetes. Upon pretreatment of stem cells with Granulocyte-Colony Stimulating Factors, injected stimulated stem cells into patients with adult onset Type 2 diabetes and adult onset neurodegenerative diseases, such as Parkinson's Disease, are able to initiate repair or to enhance the effect over Granulocyte-Colony Stimulating Factors alone. The patient provided these stern cells is also administered Granulocyte-Colony Stimulating Factors for further benefit. Alternatively, patient with the adult onset diseases is injected with stem cells that are not pretreated with Granulocyte-Colony Stimulating Factors but subsequent treatment and provided with an effective dose of Granulocyte-Colony Stimulating Factors to enhance response.

[0042] In cases where the patient has a low or depleted level of stein cells due to age, being immunocompromised due to diseased, sickened state or like condition, the preferred treatment of patients requiring supplemental doses stem cells from those donors are those individuals that are histocompatibility similar or that match having nearly identical major histocompatibility complex (MHC) such an close relative or preferably a twin. A preferred source of stem cells is to administer a dose of the Granulocyte-Colony Stimulating Factors to the patient or close relative and collect the stern cells by methods well known in the art. The more preferred source of stem cells would be stem cells collected from the individual when the individual is between 16 to 35 years of age and most preferred when the individual is between 18 to 30 years of age. Once the stem cells are collected, the stem cells may be subjected to additional rounds of Granulocyte-Colony Stimulating Factors in vitro to increase the numbers of stein cells prior to treatment of the patient. In other embodiments, the multiplied stern cells are subjected to preservation methods, freezing at low temperatures and storage at less than 50 C. to cryopreserve the cells until needed.

[0043] To optimize the production of stem cells in an individual, Granulocyte-Colony Stimulating Factors administered in conjunction with agonist and antagonist of ligand stromal derived factor-1 (SDF-1) to reduce expression of chemokine receptor CXDR4, Antagonist such as AMD3100 block the binding of SDF-1 and is known to cause rapid mobilization of CD34+ cells. With Granulocyte-Colony Stimulating Factors, AMD3100 (brand name Mozobil or Plerixafor) is effective in increasing the yield of CD334+ cells (J. Clin. Oncology, 2004, 1095-1102). Alternatively, an agonist peptide, CTCE-0021, may be more stable than natural SDF-1, which is rapidly cleaved by serum enzymes, has been reported to be an effective mobilizing agent. Other methods suggested to increase SDF-1 degradation are protease inhibitors such as MMP-9.

[0044] During the present invention, it was found that GCSF and likely related compounds are biological factors capable of mobilizing stern cells in the body and repairing or restoring specific functions of tissues. For any given cellular function, stem cells provide progenitor cells required by the body to restore specific functions of those tissues as evidenced by the effects on the more complex condition of Type 2 diabetes described herein. GCSF can cause the replication of those cells in response to specific mediators.

[0045] In another condition having a similar age related onset to Type 2 diabetes is age related androgenetic alopecia, referred to as male pattern baldness, which is the main cause of hair thinning or hair loss in adults. It is know that men with androgenic alopecia have lower levels of testosterone and increased amount of sex hormone-binding globulin that binds to testoterone. Interestingly, sex hormone-binding globulin is down regulated by insulin. Common but less than desirable treatments include minoxidil applied to the scalp to stimulate hair follicles and finasteride as an oral administration to slow hair loss but can have significant side effects. Finasteride acts to inhibit type II 5 alpha-reductase and subsequently inhibits conversion to DHT. Other routine treatments include hair transplants using plugs of hair from other areas of the scalp where hair continued to grow.

[0046] Recent investigations on improved treatments for hair loss have focused on those stem cells that are ultimately responsible for continued growth of hair. The growth of hair is not synchronized and grows in cycles in humans. It has three distinct phases of the hair growth cycle: anagen, catagen and telogen. The last phase, telogen, the keratinization is completed and the keratinized hair falls out. To replace the lost hair follicle, a new matrix is gradually formed from progenitor stem cells that are gathered in the basal layer in the outer root sheath bulge of the hair follicle. A new hair starts to grow from the matrix and the follicle returns to anagen, the hair growth phase.

[0047] Matrix cells derived from stem cells are under the influence of substances that control their growth and differentiation produced by cells of the dermal papilla. A large number of these factors are cytokines that influence or inhibit the proliferation of hair matrix cells and hair growth. Two factors believed to impact hair growth are keratinocyte growth factor (KGF) and insulin-like growth factor I (IGF-I) in animals.

[0048] Interestingly, nails are hardened epidermal variants of hair that grow continuously from the nail organ. The nail organ is an epidermal invagination on the dorsal side of the digit tips that is an equivalent of a large hair follicle. At the proximal end of the nail lies a matrix, a mass of stem cells that is equivalent to the pre-matrix cells in the hair follicle. The matrix is the immediate source of the cells that forms the nail. Unlike the loss of hair, typically nails become significantly weaker in aging as well as in response to fingernail cosmetics resulting in more brittle and flexible nails.

[0049] It has been unclear whether the loss of hair in humans, or androgenic alopecia, is caused by the inability of stem cells to mature into matrix cells that produce hair or due to a loss of stem cells or a subpopulation of stem cells that differentiate into matrix cell producing hair or a combination of both. However, a recent study suggested that balding might arise from stem cell activation rather a reduction in the number of stem cells in follicles present (J Clin Invest 2011, 121, 613). Further, they found that the same number of stem cells in the balding area, however the number of progenitor stem cells was markedly depleted in the scalp.

[0050] Other current approaches are exploring stern cell therapy for hair loss treatment. There has been considerable interest by numerous investigators in transplanting stem cells into hair follicles that are able to mature into functioning hair producing matrix cells. In one example, Hantash et al have taught of method of restoring hair by first irradiating the skin to create elongated voids in the skin. Once the voids created, stem cells such as hair follicle stern cells can be transplanted into the voids (U.S. Pat. App, Nos. 2007/0212335 and 2012/0238941). They teach that stem cells can be harvested from a variety of sources including adult hair follicles. In other related works, Cambier al taught a method to immortalize adult stem cells obtained from harvesting stem cell types of interest from the tissue or organ of interest for the treatment of human diseases (U.S. Pat. App. No. 2007/0116691). These immortalized stern cells can be used as a long term source for transplantation.

[0051] In a very recent article by Toyoshima et al (Nat. Commun. 2012, 3, 784), the authors demonstrated fully functional hair regeneration in a nude mouse model using bioengineered hair follicle germ transplanted intracutaneously. The bioengineered hair follicle germ was a bioengineered vibrissa follicle germ reconstituted with adult follicle-derived cells. This approach is very promising but transplantation requires invasive surgery to introduce the bioengineered hair follicles. It was not yet determined if these engineered follicles remain capable of producing hair in the long term. All of these studies will need to reproduced in humans to completely understand if stem cell transplantation will work in human subjects.

[0052] A new alternative topical treatment for hair loss was reported using an FDA-approved glaucoma drug, bimatoprost, that is commercially available to lengthen eyelashes may also be able to induce the growth of hair in the human scalp. Clinical trials are underway to determine if the bimatoprost is effective beyond growing and thickening hair in androgenetic alopecia where the hair follicle is deficient in progenitor matrix cells.

[0053] In a Canadian Application, CA2008/002019, related to the present invention, applicants reported that GCSF subcutaneously injected into mice at a similar dose as that used in humans, where the mice were shaved and treated with depilatory cream, showed an approximate 20% increase in hair growth as well as it was noted the hair had thicker shafts. Other factors in conjunction with GCSF promoting hair growth and hair thickening in mice included Vascular Endothelial Growth Factor and Epidermal Growth Factor. In same invention, the applicants used a model of temporary hair loss to study the effect of GCSF or other factors. Mice were injected with cyclophosphamide, a chemotherapy drug, to inhibit hair growth. It should be noted that a side effect of GCSF treatment in chemotherapy patients for neutropenia is hair loss. Surprisely, the applicants used over 10 fold higher dose rate per kilogram than used in human therapies and their previous study in patent application. In both untreated and treated mice, hair grew however the rate of hair growth in mice injected with the large dose of GCSF showed a two fold increase in hair growth compared to control. It should be noted that GCSF was given one day after injection with cyclophosphamide, suggesting that the effect would be protective. The hair growth in control mice underscores that the model does not to accurately reflect the deficient stem cells in androgenetic alopecia.

[0054] In the present invention, it was surprisingly found that GCSF was able to re-grow hair when administered in multiple consecutive courses of GCSF. Further, the hair that was restored was retained upon less frequent, routine administration of GCSF administered. In addition, GCSF was able to increase both fingernail cuticle growth and density.

EXAMPLE 1

Reversal of Symptoms with Parkinson's Disease with Granulocyte-Colony Stimulating Factor

[0055] The patient was a 61 year-old male with a six year history of Parkinson's Disease. When first diagnosed two years prior, the patient was started on 100 milligrams Amantidine twice daily and 5 milligrams Selegiline twice daily and on increasing doses of Permax, 25 micrograms to 250 micrograms three times daily.

[0056] A history taken immediately before starting the dosing of Granulocyte-Colony Stimulating Factor (GCSF) was typical of adult onset Parkinson's disease. The patient had blood work performed 7 days before treatment which included CBC, ESR, urine analysis and chemistry screen. In summary, the male patient, approximately 67 kg body weight diagnosed with Parkinson's Disease had history of modest hyperlipidemia which was controlled on medications. Physical exam was normal except for some cogwheeling especially on the right. Granulocyte-Colony Stimulating Factor was started at a daily dose of 330 micrograms (1.1 ml) through injection for 5 consecutive days. Patient experienced slight bone pain on second and third days of therapy. About 3 weeks after Granulocyte-Colony Stimulating Factor therapy was started, patient realized that he could smell, something that he had not been able to do for more than 5 years. This observation is consistent with improvement in olfactory nerve function. About a week later, the patient found that the orthostatic hypotension that had given him trouble for the last 2-3 years and had required him to wear support stockings, had disappeared. In addition, the patient was able to stand up quickly from a sitting position or walk up stairs without becoming severely light-headed. The patient's handwriting, previously totally unreadable, was greatly improved and was legible to others. It was observed by his wife that the patient had an improved facial affect. After about two months, a plateau was reached.

[0057] Surprisingly, it was found that Granulocyte-Colony Stimulating Factor treatment of a patient with Parkinson's disease was followed four weeks later by quantifiable evidence of efficacy utilizing symptoms that indicated an effect on both the Central Nervous System and Peripheral Nervous System (PNS). The patient improved in several CNS symptoms that included resting tremor, rigidity, postural instability, and micrographia. Four accepted PNS symptoms of Parkinson's disease, the loss of sense of smell, orthostatic hypotension and urinary and gastrointestinal incontinence disappeared during the third and fourth weeks following the start of therapy. The disappearance of orthostatic hypotension was not complete but was greatly reduced. Unexpectedly, GCSF caused reversal of one or more symptoms of Parkinson's Disease upon treatment with the first dose. The reversed symptoms included orthostatic hypotension, resting tremor, rigidity, postural instability, handwriting and urinary and gastrointestinal incontinence.

EXAMPLE 2

Long Term Treatment of Parkinson's Disease Using Granulocyte-Colony Stimulating Factor

[0058] The long term treatment of Parkinson's Disease by GCSF was followed using a standardized system approved by neurologists, the Unified Parkinson's Disease Scale (UPDS), to more completely access the efficacy of treatment, UPDS utilizes over 40 distinct clinical evaluations that are predominantly based mostly on CNS symptoms. Analysis was performed approximately 2 to 4 weeks after injection of the first dose. To more fully understand the effect of the GCSF, the data was analyzed to determine whether a correlation existed between dose of GCSF and reversal of symptom score. The patient received injection of the same dose of GCSF every six to eight weeks and was periodically evaluated by UPDS criteria. In FIG. 1, the score of UPDS was plotted over the course of two and one-half years. The correlation coefficient was 0.93 suggesting the reversal was progressive upon GCSF treatment and was statistically significant (P=0.001). The final score obtained by the patient approached the score that a non-patient might receive.

[0059] Because UPDS evaluates mostly CNS symptoms of Parkinson's Disease, four indicators of PNS symptoms were also evaluated included urinary and gastrointestinal incontinence, orthostatic hypotension and loss of smell. The induction of orthostatic hypotension was evaluated by taking blood pressure before and after going from a lying down position to sitting and then standing position and each time taking blood pressure with an automatic apparatus. A drop of 50 mm was associated with developing lightheadedness and orthostatic hypotension. All four indicators were reversed relative to the state prior to GCSF treatment. In fact, the patient was able to exercise without fainting or having to rest (orthostatic hypotension), enjoyed foods and wine (smell), resolved urinary and gastrointestinal incontinence, and gained sense of smell resulting in a higher quality of life.

EXAMPLE 3

Reduction of Blood Glucose Levels in Type 2 Diabetics Administered with Granulocyte-Colony Stimulating Factor

[0060] Other benefits were observed upon treatment with Granulocyte-Colony Stimulating Factor in the same patient as described in Example 1. The patient exhibited a two year history of slightly elevated blood glucose determinations (average fasting blood sugar ranging up to 115 mg/Dl). Patient had a slightly elevated hemoglobin A1C (6.5 versus normal of <6.0. He was diagnosed as Type 2 Diabetes and was recommended to a Diabetics clinic for classes on diet and exercise for dealing with this disease. Although dieting and exercise provided some reducing it was not solely sufficient in controlling blood glucose levels below the 110 mg/dl.

[0061] After treatment with Granulocyte-Colony Stimulating Factor it was observed that the blood sugar averaged 104 mg/Dl for 15 specimens before treatment and decreased to 92 mg/Dl during the period starting three weeks after the start of drug (and at the same time that Parkinson's symptoms disappeared. The difference was highly significant (P=0.001).

EXAMPLE 4

Long Term Controlled Reduction of Blood Glucose Levels in Type 2 Diabetics Administered with Granulocyte-Colony Stimulating Factor

[0062] To examine the long term effect of Granulocyte-Colony Stimulating Factor on a patient with adult onset Type 2 diabetes, the patient in Example 2 was monitored for approximately two and one-half years for fasting blood glucose almost every morning. Starting with the first day of therapy with GCSF, every 5 day period was averaged and the mean used to compare blood glucose levels. FIG. 3 shows the analyzed data from blood sugar determinations around six courses of GCSF. Each time after the GCSF is administered the blood sugar dropped well within the normal range after about twenty to thirty days after the start of therapy. In one case drug was not administered until late in the period of follow-up and then the nadir was reached about twenty-five days after the start of that course. All curves reached a nadir ranging between 20 and 30 days after the start of therapy. Most blood glucose determinations remained below 110 mg/Dl for about 2 months after treatment. It was unexpected that a drug, and more surprising GCSF, is able to reduce blood glucose for extended periods beyond one day, even more surprising one week and most surprising one month. The patient was also able to ingest limited amounts of Dove bars without any problems. GCSF provides a consistent and more desirable approach to regulate blood glucose levels in Type 2 diabetics with less potential side effects of multiple mechanisms employed to control Type 2 diabetes beyond other drugs.

EXAMPLE 5

GCSF Supplemental Production of Stem Cells in a Close Donor or Self to Improve Control in the Reduction of Blood Glucose Levels in Type 2 Diabetics

[0063] An individual was treated with stem cells obtained from self or a close relative as a way to increase the effect of GCSF in controlling on blood glucose levels. In the later case, an identical twin was examined for MHC profile and found to be a match. To produce a supplemental amount of stem cells in the identical twin, the twin was administered Granulocyte-Colony Stimulating Factor a daily dose of 330 micrograms through injection for 5 consecutive days. Patient experienced slight bone pain on second and third days of therapy. After 7-10 days, the blood of the patient was subjected to leukaperesis to collect peripheral blood stem cells from the mobilized peripheral blood progenitors, Preferably, CD34+ stern cells are isolated from peripheral blood stem cells and HLA matched donors.

[0064] The stem cells obtained from a Neupogen-treated twin were injected into the patient with Type 2 diabetes that was previously treated with GCSF. After 1-2 weeks, the patient was administered a course of GCSF; 330 micrograms (1.1 ml) through injection for 5 consecutive days. It was found that the level of blood glucose decreased significantly from the prior treatment and remained lower for a longer period that the prior treatment.

[0065] The exogenous treatment using isolated stem cells of the Type 2 diabetic patient was also repeated with the patient's own stem cells, however the increase was less than that observed with treatment of stem obtained from the identical twin. Preferably, self-produced stem are harvest from the individual when the individual is a young adult between the ages of 16 to 35 years of age. The optimal ages are between 18 and 30 years of age. The harvested stem cells are subjected to preservation methods, such as cryopreservation until needed.

EXAMPLE 6

GCSF Acceleration of Hair Growth and Fingernail Growth and Density

[0066] Yet other benefits of the Granulocyte-Colony Stimulating Factor treatment observed was the re-growth of hair in a human patient with androgenic alopecia.

[0067] To re-grow of hair in the balding scalp, the patient was treated with an increased frequency of the courses of Neupogen administered. Prior to treatment, the patient had received courses of GCSF as described in Example 2 and Example 4, however no significant re-growth of hair was observed. Prior to the increased frequency of treatment, the patient exhibited moderate hair loss and thinning, corresponding to a rating of 5 on Hamilton-Norwood Scale for hair loss, ranging from 0 for no hair loss to 7 being complete hair scale loss. Neupogen (480 ug; 5-6 ug/kg body weight) was administered for each day for four days followed by a period of no treatment for four days. This regimen was repeated for a total of six times consecutively. After the patient continued on their normal periodic treatment of GCSF, being administered with GCSF courses every 6-18 weeks. Surprisingly, about 4-6 months after GCSF treatment, the substantial growth of new hair, or re-growth of hair, on the subject's scalp resulted in a improvement of hair growth to 2 on the Hamilton-Norwood Scale. The hair was similar in texture and distribution to early stages of male pattern baldness in the individual. In dramatic comparison to the patient's identical twin, as shown in FIG. 5, the untreated twin brother had a rating of 6 on the Hamilton-Norwood Scale. Both individuals had a similar amount of hair loss prior to administration of GCSF to the treated patient.

[0068] After the increased courses were completed, the administration of Neuprogen returned to a treatment of every 6-18 weeks. It was observed the hair grew much more quickly in the months following administration of the increased courses. The rate of hair growth and density has been maintained the same growth since the patient treatment has returned to routine courses during the past 2 years. These findings showed that once the amount of desired hair growth is obtained, the lower frequency rate is suitable to retain hair growth.

[0069] For consecutive courses, GCSF is administered for about 3 to 6 days and the treatment is repeated about every 3 to 8 days for two or more consecutive courses, preferably 3 to 6 courses. The administration of effective consecutive courses and periodic courses of GCSF is administered at a daily dose rate of 0.1 ug to 100 ug, preferably 0.1 to 10 ug per kg, more preferably 0.1 to 5 ug per kg and most preferably 0.1 to 2 ug per kg body weight.

[0070] Other factors can be used to supplement hair growth and density with GCSF administration. For example, based on bimaprost findings stimulating hair growth and thickening, bimaprost, and like compounds working on related receptors in the follicle area, may be effective to increase hair volume once the damaged progenitor stem cells have been replenished. Other factors that may enhance include Vascular Endothelial Growth Factor and Epidermal Growth Factor reported earlier to stimulate hair growth. GCSF treatment would also be effective as a companion treatment for stem cell transplantation either prior to, after or both, including pre-conditioning of the stem cells with GCSF prior to transplantation,

[0071] This is the first report where GCSF was demonstrated to re-store hair growth in a human patient with androgenic alopecia. The results showed that GCSF was able to restore hair matrix cells functionality to growth hair and further treatments could obtain additional hair growth. The consecutive course regimen of GCSF not only mobilizes progenitor stem cells but also facilitates their maturation into hair producing matrix cells. It is surprising that many references have suggested that hair loss may occur with GCSF treatment, however this may be complicated by the fact that GCSF in limited courses in chemotherapy.

[0072] Because the hair matrix cells are similar to fingernail matrix cells, it was also observed that the cuticle density and growth improved dramatically during the same period. Prior to treatment, the fingernails of patient were very flexible and had a slow growth rate as well as the nails were often chipped. In the same subject treated with the multiple courses of GCSF administration described for hair re-growth, it was found that the fingernails had become much denser and grew at a faster rate similar to that observed for their hair, Identical effective consecutive courses and periodic courses of GCSF for hair would used be for cuticle, fingernail and toe nail GCSF treatments.

Other Embodiments

[0073] The description of the specific embodiments of the invention is presented for the purposes of illustration. It is not intended to be exhaustive nor to limit the scope of the invention to the specific forms described herein. Although the invention has been described with reference to several embodiments, it will be understood by one of ordinary skill in the art that various modifications can be made without departing from the spirit and the scope of the invention, as set forth in the claims. All patents, patent applications, and publications referenced herein are hereby incorporated by reference. Other embodiments are within the claims.