Biomarker Assessments of Leukine During Treatment of Parkinson's Disease

Primary Objectives: There are three study goals. First, the investigators will determine the
safety of a 6 month regimen of Leukine administered as a weight-based dose at 3 µg/kg/day for
5 days (week), followed by a 2-day holiday (weekend). This 6 month (n=5) pilot study will
extend the prior 2 month observation tests towards the goal of assessing the safety of
Leukine for treatment of Parkinson's disease (PD). Clinical signs and symptoms will be
measured by personal well-being, physical, and neurological examinations (UPDRS Parts I, II,
III, and IV assessments) and blood tests (CBC with differential, total T cell count, and a
comprehensive metabolic sera panel). Second, the investigators will assess regimen
tolerability administered in a dose reduction, from 6 µg/kg/day without interruption, to 3
µg/kg/day with 2 day drug holidays. The lowered dose was chosen based on known tolerability
and parallel-linked immune reconstitution seen in cancer-associated disease treatments. Due
to fragility of the patient population and prior recorded adverse events the proposed dose
reductions are justified.

Secondary Objectives:

Over a course of 6 months, the effects of treatment on defined adaptive immune deficits in PD
as measured by analysis of peripheral blood mononuclear cells collected before, during, and
after cessation of Leukine administration will be examined. Individual T cell parameters that
include links between T cell function and subset analyses and clinical neurological signs and
symptoms will be examined. These immune parameters will be serially examined as they may
contribute to the immune deficits in PD. Thus, timed analyses of changes in T cell phenotypes
and/or function will be completed. In addition, the investigators will assess the functional
stability of the immune deficits in PD and will determine whether the immune deficits of PD
are consistent during baseline data collection. The potential Leukine-induced motor control
and mobility improvements will be determined by assessing UPDRS part I, II, III, and IV
scores off treatment and on treatment. Specifically, over the course of this six-month
treatment study, various biomarkers will be assessed including: T cell markers of immune
activation, DNA methylation status, and B cell and bone marrow progenitor cell markers. We
hope to uncover the time course of Treg induction, as well as link the pharmacokinetics of
Leukine treatment (not previously recorded) to changes in specific biomarkers. Additionally,
changes in the humoral response following extended Leukine treatment will be assessed by
measuring the presence of antibodies against Leukine and alterations in B cell frequencies.

Inclusion Criteria:

- 1. Onset of bradykinesia and 1 or both of the following: rest tremor and/or rigidity

2. Asymmetric onset of clinical signs

3. Progressive motor symptoms

4. Age at onset 35-85 years

5. Duration of PD symptoms of at least 3 years

6. Female subjects must be either:

1. Not pregnant, not breastfeeding, and not planning on becoming pregnant during the
study;

2. Not of childbearing potential, defined as one who has been postmenopausal for at
least 1 year and with follicle stimulating hormone (FSH) levels in the laboratory
defined postmenopausal range, or has been surgically sterilized, or has had a
hysterectomy at least 3 months prior to the start of this trial; or

3. If of childbearing potential, must agree to use an effective method of avoiding
pregnancy to the end of the trial and must have a negative serum beta-human
chorionic gonadotropin (β-HCG) test. Effective methods of avoiding pregnancy are
contraceptive methods used consistently and correctly (including implantable
contraceptives, injectable contraceptives, oral contraceptives, transdermal
contraceptives, intrauterine devices, diaphragm with spermicide, male or female
condoms with spermicide, or cervical cap), abstinence, or a sterile sexual
partner.

7. Must be stage 4 or less according to the Hoehn and Yahr scale

Exclusion Criteria:

- 1. Atypical features indicative of a Parkinson-Plus disorder (Progressive Supranuclear
Palsy (PSP), Multiple System Atrophy (MSA), Corticobasal Degeneration (CBD)) including
cerebellar signs, supranuclear gaze palsy, apraxia and other cortical signs, or
prominent autonomic failure

2. Neuroleptic treatment at time of onset of parkinsonism

3. Active treatment with a neuroleptic at time of study entry

4. History of repeated strokes with stepwise progression of parkinsonism

5. History of repeated head injury

6. History of definite encephalitis

7. More than one blood relative diagnosed with PD

8. Prominent gait imbalance early in the course (< 5 years)

9. Mini-mental state examination score <26

10. Hematological malignancy or coagulopathy

11. Abnormal blood analyses: hematocrit <30; WBC>11.5; clinically significant
laboratory data (e.g. alanine aminotransferase [ALT] or aspartate aminotransferase
[AST] 3x the upper limit of normal [ULN]), or any abnormal laboratory value that could
interfere with the assessment of safety in the judgment of the investigator;
significant abnormalities on the clinical examination, vital signs, and clinical
chemistry or hematology results (excluding findings of Parkinson's disease), that may
interfere with the study or present a safety risk for the subject as judged by the
clinical investigator charged in the care of study participants

12. Serious medical illness or co-morbidity that may interfere with participation in
the study

13. Brain surgery for parkinsonism (DBS, cell implantation, gene therapy)

14. History of an autoimmune disorder or systemic inflammatory disorder deemed
significant by physician

15. Immunostimulatory or immunosuppressive treatment (including amphet-amines or
systemic corticosteroids) within 90 days

16. Exclusively unilateral parkinsonism for longer than 3 years

17. Known hypersensitivity to GM-CSF, yeast-derived products

18. Current lithium treatment

19. Individuals with current diagnoses of alcohol or substance abuse/dependence

20. Anyone who is not appropriate for participation in this research protocol as
deemed by the principal or co-investigator

21. Anyone who has previously been treated with GM-CSF as an immunomodulatory therapy

22. Anyone with poor venous access