The P65Oncofetal Protein as a Novel Tumor Marker in Detection of
Breast, Prostrate and Other Cancers CSU 5-s6904
Amendment No. 1
Research Agreement
between
The Board of Governors of the Colorado State University System
acting by and through Colorado State University
and
Xpention Genetics
This Amendment is made and entered into by and between Xpention Genetics
hereinafter referred to as the SPONSOR, and the Board of Governors of the
Colorado State University System, acting by and through Colorado State
University, hereinafter referred to as UNIVERSITY.
NOW THEREFORE,
The parties mutually agree to the following revisions to this Research
Agreement:
Article 1 - Scope of Work The University agrees to perform for the Sponsor the
additional research described in the Scope of Work, Exhibit A1 attached hereto
and incorporated by this reference, under the direction and supervision of the
Principal Investigator, Dr. Xxxxx Xxxx, in accordance with any service
milestones or periodic deliverables specified on Scope of Work Attachment.
Article 2 - Term
This Agreement is extended to November 1, 2006
Article 3 - Payment The Sponsor agrees to pay the University for research
performed under this Agreement in a fixed price amount as set forth in the
Budget Attachment, Exhibit B1 attached hereto and incorporated by reference. The
University reserves the right to reallocate funds between approved budget
categories. Payment will be made in accordance with the schedule provided below:
Fifty percent (50%) ($7,665) upon acceptance of agreement; Forty
percent (40%)($6,132) mid-way through project (date: May 1, 2006); Ten
percent (10%) ($1,533) upon submission of final report.
All other terms and conditions of this agreement remain the same.
IN WITNESS WHEREOF, the parties hereto have caused this Agreement to be executed
as of the date set forth herein by their duly authorized representatives.
The Board of Governors of the Xpention Genetics
Colorado State University System,
acting by and through Colorado
State University
/s/ Xxxx ???? /s/ Xxxxx Xxxxxxxx
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Director, ??? Programs 11/2/05 President 10/26/05
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Title Date Title Date
EXHIBIT A1
Description of Project
Specific Aims
Aim of this project is the development of a molecular test for early detection
of cancer in dogs. This test will target a genetic marker which levels are
greatly increased in the early stages of cancer development. Using "Real Time
Fluorogenic PCR (F-PCR)"technology, this test will accurately measure level of "
P65 gene expression" during the different clinical stages of cancer development.
By comparing levels of P65 before, during and after therapy it will be possible
to determine 1) rate of response to cancer treatment 2) prognosis outlook.
Several studies using other genetic markers have shown that these two parameters
are of paramount importance in deciding whether a specific chemotherapy protocol
should be continue or terminate because a poor long term outlook.
Technology Background
"Real Time Fluorogenic PCR" assay for P65
Previous studies performed in canine lymph sarcoma showed that during early
stages of tumor development level of P65 in the blood of affected animals
increases up to 50 folds. These studies also showed that during the course of
chemotherapy levels of P65 return to normal if the animal successfully responds
to the treatment. Techniques used during these studies are based on the use of
immunological assays. Several limitation exist when such techniques are
employed: 1) low sensitivity in detecting P65 levels; 2) lack of correlation
between the presence of antibodies in serum and stages of tumor progression; 3)
antibodies are detected at a later stage of tumor progression when compare to
changes in genetic levels of the target gene. F- PCR overcomes the above
limitations making it possible to perform much earlier diagnosis of cancer.
F-PCR is the most sensitive technology available to date to detect and
quantitate genetic levels in a variety of tissues. F-PCR will be employed to
develop a clinical assay for P65 detection in blood cells of dogs lymphosarcoma
and other type of cancer. This is the first cancer test that will be able to
detect the presence of a genetic cancer marker in circulating cells rather than
tissues. The implications of this approach are of paramount importance:1)
eliminates the need of invasive procedures such biopsies or explorative
surgery;2) allows more frequent assay screenings cutting waiting time; 3)
reduces diagnostics errors often present when histopathology is performed; 4)
allows correct correlation between tumor stage and chemotherapy response and 5)
allows faster determination of rate of success to chemotherapy.
Assay Development
We anticipate a 12 month period in order to complete the development of the P65
molecular assay for detection of cancer in dogs. Listed below is a description
of the developmental phases
Phase I
- Development of RNA extraction protocol from canine blood samples
- Designing of the P65 oligonucleotides primers and FAM probe
- Designing of GAPDH oligonucleotides primers and VIC probe
Phase II
- Endogenous control calibration
- Baseline determination of P65 expression
- Delta Ct correlation
Phase III
- Multiplex assay development
Phase IV
- Assay validation
Scope of work to be completed by Animal Cancer Center, Colorado State University
under the supervision of Dr Xxxxx Xxxx. Blood samples will be obtained from 60
canine patients with cancer at the time of initial diagnosis. Informed consent
will be obtained from each pet owner as well as ACUC approval. A variety of
tumor types will be represented as agreed upon with the sponsor. Samples will be
collected into specified tubes (provided by the sponsor) for RNA preservation
and will be processed according to procedures provided. Samples will be frozen
and stored and sent in batches to the sponsor. The time required to complete
this portion of the project is dependant on the clinical case load and the
tumors types selected by the sponsor. It is anticipated 6 to 10 months will be
required.
Budget justification
Attached (appendix 1) is a budget for 60 samples based on a per sample charge.
This charge includes supplies, personnel, and other costs associated with blood
collection, processing and storage.
EXHIBIT B1
Budget for molecular detection of p65 in canine blood samples
Sample cost
$175 per sample, 60 samples requested $10,500
Indirect cost
Non clinical trial rate at CSU 46% $4830
Total $15,330
