Trial of Bi-shRNA-furin and Granulocyte Macrophage Colony Stimulating Factor (GMCSF) Augmented Autologous Tumor Cell Vaccine for Advanced Cancer

Preliminary studies with a variety of vaccines suggest target accessibility (potential
immunogenicity) in a variety of solid tumors to immune directed approaches. In an effort to
overcome limitations of immunostimulatory cancer vaccines, we have designed a novel
autologous vaccine to address inability to fully identify cancer associated antigens, antigen
recognition by the immune system (i.e. antigen to immunogen), effector potency, and
cancer-induced resistance. We have completed clinical investigations using two different gene
vaccine approaches to induce enhancement of tumor antigen recognition which have demonstrated
therapeutic efficacy. Specifically, both the use of a GMCSF gene transduced vaccine (GVAX®)
and a TGFβ2 antisense gene vaccine (Lucanix®), in separate trials, have demonstrated similar
beneficial effects without any evidence of significant toxicity in advanced cancer patients.
The GMCSF transgene directly stimulates increased expression of tumor antigen(s) and enhances
dendritic cell migration to the vaccination site. TGFβ2 blockade following intracellular
TGFβ2 antisense gene expression reduces production of immune inhibiting activity at the
vaccine site. This appears to be one of the primary mechanisms of inhibition of immune
responsiveness in glioblastoma and lung cancer. In a subsequent Phase I trial we combined
both active principles in one autologous vaccine, TAG. TAG vaccine has an excellent safety
profile in the first nineteen patients treated (enrollment open to any solid tumor) with one
documented CR (melanoma). However, TGFβ1 is the dominant TGFβ family inhibitory effector in
the majority of other solid tumors. We describe a unique method of inhibiting both TGFβ1 and
TGFβ2 through RNA interference with Furin. We will harvest autologous cancer cells from
patients with advanced refractory cancer. We have constructed a bi-shRNAfurin / GMCSF
(Vigil™) expression vector plasmid and have successfully demonstrated preclinical activity of
the vector function following transfection by electroporation and irradiation of ex vivo
autologous tumor cells.

Tissue Procurement Inclusion Criteria:

1. Presumptive or histologically confirmed advanced or metastatic non-curable solid tumor
(if limited to a single lesion, and may not be a candidate for curative surgery or
radiation therapy).

2. For the purpose of the Pediatric study patients with histologic diagnosis of ESFT
including: Ewing's sarcoma or primitive neuroectodermal tumor (malignant
neuroepithelioma) of the bone or soft tissues, Askin's tumor of the chest and with
central nervous system tumors are eligible.

3. Patients with recurrent or refractory ESFT. Patients with de novo poor prognosis/high
risk ESFT: (Eligible for vaccine manufacturing at diagnosis but ONLY ELIGIBLE FOR
IMMUNOTHERAPY IF DEMONSTRATES PERSISTENT/RECURRENT/ REFRACTORY DISEASE)

1. Large tumors > 8 cm

2. Pelvic osseous tumors ANY SIZE

3. Bilateral pulmonary metastasis

4. >2 unilateral pulmonary metastasis

4. Clinically (medically) indicated procedure (i.e. biopsy of lesions of recurrent
disease, palliative management via resection, thoracentesis, etc.) to collect tumor in
sufficient quantity ("golf ball size" estimated weight 10-30 grams, pleural, and/or
ascites fluid estimated volume ≥ 500 mL) for vaccine processing.

5. For ESFT patients age ≥12 years.

6. Age ≥18 years (non-ESFT candidates) ECOG performance status (PS) 0-1. Pediatric
patients with Lansky or Karnofsky Performance Status Scale ≥ of 50%.

7. Estimated >4 months survival probability.

8. Ability to understand and the willingness to sign a written informed consent document.
Pediatric patients must sign an assent with a parent or legal guardian sign a written
informed consent.

Inclusion Criteria:

1. Histologically confirmed advanced or metastatic non-curable solid tumor (if limited to
a single lesion may not be a candidate for curative surgery or radiation therapy).
Successful vaccine manufacture has resulted from tissue/fluid obtained from the
following major organ systems: digestive, endocrine, reproductive, respiratory, and
urinary.Individuals manufactured under CL-PTL 105 (Phase II Ovarian) may be eligible
for enrollment without advanced or metastatic disease.

2. Patients with well differentiated thyroid cancer are eligible for protocol as follows:

1. Surgically unresectable locally recurrent disease and/or metastatic disease
following RAI ablation (if locally recurrent and ultrasound (US) positive,
baseline FDG-PET or MRI will be obtained).

2. Patients with microscopic and/or gross extra thyroidal disease extension without
RAI uptake but with a) FDG-PET positive disease or b) suppressed thyroglobulin >1
ng/mL or c) stimulated thyroglobulin >10 ng/L.

3. Patients with tracheal/esophageal involvement. High mitotic activity or necrosis
in pathology does not exclude from the study.

Note: in Categories a and b, patients can be followed using US locally in addition to
standard diagnostic followup menu but, if US only is positive, a FDG-PET or MRI will
be obtained. If negative, a rising thyroglobulin titer is required in which case
response will be monitored by continued US and suppressed and/or stimulated
thyroglobulin. Thyroglobulin titer cannot be used if anti-thyroglobulin antibodies are
present).

3. Completed all acceptable therapies with curative intent that are the current standard
of care for their respective diseases. If no conventional therapy available, patient
may participate after review by sponsor.

4. Recovered from all clinically relevant toxicities related to prior therapies

5. Patients will be allowed to participate following single prior CNS treatment with
stereotactic radiotherapy whole brain irradiation and stable without steroid
requirement for ≥2 months or following ≥2 prior CNS treatments with stereotactic
radiotherapy whole brain irradiation and stable without steroid requirement for ≥4
months.

6. For ESFT patients age ≥12 years.

7. Age ≥18 years (non-ESFT candidates)

8. ECOG performance status (PS) 0-1.Pediatric patients with Lansky or Karnofsky
Performance Status Scale ≥ of 50%.

9. Estimated >4 month survival probability.

10. Normal organ and marrow function as defined below:

Absolute granulocyte count ≥ 1,500/mm3 Absolute lymphocyte count ≥ 500/mm3 Platelets ≥
100,000/mm3 Total bilirubin ≤2 mg/dL AST(SGOT)/ALT(SGPT) <2x institutional upper limit
of normal Creatinine <1.5 mg/dL

11. Ability to understand and the willingness to sign a written informed consent document.
Pediatric patients must sign an assent with a parent or legal guardian sign a written
informed consent.

12. Negative pregnancy test.

13. Patients must be off all "statin" drugs for ≥2 weeks prior to initiation of therapy.

Exclusion Criteria:

1. Surgery involving general anesthesia, radiotherapy, steroid therapy, or immunotherapy
within 4 weeks prior to vaccination. Chemotherapy within 3 weeks prior to entering the
study. Palliative radiotherapy is allowable. Collection of lumenal tissue for vaccine
manufacture must be avoided.

2. Patients must not have received any other investigational agents within 30 days prior
to vaccination.

3. Concurrent tumor-specific hormonal therapy or antiestrogens. (Individuals manufactured
under CL-PTL 105 (Phase II Ovarian) are not subject to this exclusion).

4. Patients with known active or symptomatic brain metastases prior to vaccination.

5. Patients with compromised pulmonary disease.

6. Short term (<30 days) concurrent systemic steroids ≤0.25 mg/kg prednisone per day
(maximum 7.5 mg/day) and bronchodilators (inhaled steroids) are permitted; other
steroid regimens and/or immunosuppressives are excluded while receiving vaccine.
Patients requiring steroids following previous CNS radiation for metastatic disease
are excluded.

7. Prior splenectomy unless Howell-Jolly bodies are absent.

8. Prior malignancy (excluding nonmelanoma carcinomas of the skin) unless in remission
for 2 years.

9. Kaposi's Sarcoma.

10. Uncontrolled intercurrent illness including, but not limited to ongoing or active
infection, symptomatic congestive heart failure, unstable angina pectoris, cardiac
arrhythmia, or psychiatric illness/social situations that would limit compliance with
study requirements.

11. Patients who are pregnant or nursing.

12. Patients with known HIV.

13. Patients with chronic Hepatitis B and C infection. For patients with hepatocellular
carcinoma (HCC), the presence of chronic HBV and HCV is NOT an exclusion. Patients
must have a viral titer <50 IU/ml x 2 at a minimum of 2 weeks apart.

14. Patients with uncontrolled autoimmune diseases.