For IBC Use Only
Protocol Number:
Received:
Approved:
IBC
(INSTITUTIONAL BIOSAFETY COMMITTEE)
PROTOCOL SUBMISSION FORM
IBC Office, 1430 Tulane Ave. TW-5, New Orleans, LA 70112
504.988.0300 (ph) 504.988.0370 (fax)
Submit the completed application by email to ibc@tulane.edu with “IBC-Application” in the
subject line.
TITLE AND PERSONNEL
Title of the study: Enhancing hormone therapy and chemo-therapy of human prostate cancer
and breast cancer
Submission Date:
Is this a new submission? YES
Principal investigator: Yan Dong
Department: Structural and Cellular Biology
Position/title: Associate Professor
School: Shool of Medicine
Email: ydong@tulane.edu
Phone: 504-988-4761
Co-investigator’s name:
Title:
Emergency phone:
Email:
Phone:
TYPE OF RESEARCH
The research described in this application involves: (select all that apply)
1. Recombinant DNA/RNA
YES
If “YES” complete Sections A, B, E and F of this application
NO
2. Infectious/Pathogenic microorganisms or toxins
YES
If “YES” complete Sections A , C, E and F of this application
NO
3. Select Agents (see appendix 1)
YES
If “YES” complete Sections A, C, D, E and F of this application
4. Animals
YES
NO
NO
If “YES” provide IACUC approval number:and 4154
5. Humans
YES
NO
If “YES” provide IRB approval number:
IBC Protocol Form- Jan/2010
Page 1 of 14
SECTION A: GENERAL INFORMATION
PERFORMANCE SITES
Research location(s)--check all that apply:
Tulane University St. Charles Campus
Tulane University School of Medicine
Tulane University School of Public Health and Tropical Medicine
Tulane National Primate Research Center
Other location(s). Specify :
Building and Room Number(s) where research will be conducted (provide buildings/room numbers
for all experiments detailed in this application), and biocontainment level of that room.
Biocontainment
Building:
Room No:
level for this
Experiment (ie. in vitro work, animal infection)
laboratory*
JBJ
472
JBJ
JBJ
462
Vivarium
BSL2
BSL2
BSL2-N (animals)
in vitro work
in vitro work
in vivo animal xenograft
Does this project involve work to be done by collaborators at other institutions?
Name of the institution(s):
If “Yes”, this institution granted approvals for:
IBC
IACUC
IRB
Don’t know
Yes
No
Yes
No
SECTION A
PROJECT OVERVIEW
Provide a description of the work to be conducted in this project. Briefly explain in language
understandable to a layperson the aim of this research and its importance to human or animal
health, the advancement of knowledge, or the benefit to society. Succinctly describe in
chronological order the experiments that will be conducted.
Our aim is to develop agents with low toxicity that can enhance the efficacy of hormone therapy
and/or chemo-therapy in human prostate and breast cancer. We plan to investigate the ability
and mechanism of these agants in enhancing the efficacy of current treatment methods. The
study could lead to the development of an effective, mechanism-driven strategy to significantly
improve the therapeutic outcome and ultimately to reduce prostate/breast cancer mortality.
IBC Protocol Form- Jan/2010
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SECTION B
Recombinant DNA Research Questionnaire.
(Check 'Yes' or 'No' to the following questions (refer to the NIH Guidelines* for more
information). The relevant section of the NIH Guidelines is referenced for each question. For
any items checked 'YES', include a thorough description of the work in the following page
(question 1).
1. Does your project include deliberate transfer of a drug resistance trait to
microorganisms that are not known to acquire the trait naturally (Section IIIA*)?
1a. If “YES”, could such a transfer compromise the use of the drug to control
disease agents in humans, veterinary medicine, or agriculture?
2. Does your project include cloning toxin molecules with an LD50 of less
than 100 nanograms per kilogram body weight (Section III-B*)?
3. Does your project include experiments involving the deliberate transfer of
recombinant DNA, or DNA or RNA derived from recombinant DNA, into
one or more human research participants (Section III-C*)?
4. Does your project include experiments using Risk Group 2, Risk Group 3,
Risk Group 4, or Select Agents as host-vector systems (Section III-D-1*)?
5. Does your project include experiments in which DNA from Risk Group 2,
Risk Group 3, Risk Group 4 , or Restricted Agents is cloned into
nonpathogenic prokaryotic or lower eukaryotic host-vector systems (Section
III-D-2*)?
6. Does your project include experiments involving the use of infectious DNA
or RNA viruses or defective DNA or RNA viruses in the presence of helper
virus in tissue culture systems (Section III-D-3*)?
7. Does your project include experiments involving whole animals in which the
animal’s genome has been altered by introduction of DNA into the germ line
(i.e. transgenic animals) (Section III-D-4, III-E-3*)
8. Does your project include experiments involving viable rDNA-modified
microorganisms tested on animals (Section III-D-4, III-E-3*)?
9. Does your project include experiments involving genetically engineered
whole plants (Section III-D-5, III-E-2*)?
10. Does your project include experiments involving more than 10 liters of
culture (Section III-D-6*)?
11. Does your project include experiments involving the formation of
recombinant DNA molecules containing two-thirds or more of the genome of
any eukaryotic virus (Section III-E-1*)?
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
* The NIH Guidelines are found in this link: http://oba.od.nih.gov/rdna/nih_guidelines_oba.html
Updated information about required levels of Biosafety are found in the
CDC/NIH Biosafety in Microbiological and Biomedical Laboratories (BMBL)
http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm
IBC Protocol Form- Jan/2010
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IBC Protocol Form- Jan/2010
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SECTION B
Recombinant DNA Research Questions
1. Address any items checked ‘Yes’ in the Recombinant DNA Research Questionnaire
(include the question number).
Question 1: To develop stable transfectants of aromatase expressing MCF-7 cells, we need
to transfect a plasmid containing zeocin resistant cassette to be used as a selective marker.
Qusetion 6: We need to use retroviral and lentiviral vectors for our eukaryotic infection to
obtain the high efficiency of transient transfection.
2. Describe any use of animals or human subjects. The approval of your protocol will not
be effective until required IACUC and/or IRB approvals have been received.
Groups of Balb/c male or female mice will be inoculated with ~5x10^6 human prostate
cancer cells or breast cancer cells to setup xenograft model. At the end point, the mice will
be euthanized and blood and tissue samples will be harvested for further analysis.
3. Identify the host(s) to be used (e.g., the target of gene transfer). Examples: E. coli, S.
cerevisiae, human/animal cells, whole animals, humans. Provide species designations for all
organisms where possible.
The host used for prokaryotic transformation is E. coli. The hosts used for eukaryotic
transfection are human prostate cancer cell lines and breast cancer cell lines.
4. Identify the vector(s) to be used. Examples: Bacterial plasmids, yeast vectors,
mammalian cell vectors, baculoviruses, transforming viruses, etc. Provide name and
description of the vector, including antibiotic resistance genes.
The vectors to be used are:
1) pCI-Neo from Promega with ampicilin resistance; 2) pGL4.19 from Promega with
ampicilin resistance;
5. Identify the nature of the insert DNA sequence, including the species of origin (i.e.,
specific gene, promoter, expressed product and function (if known). If available attach a
vector and/or insert map.
The insert DNA sequences include: 1) full length of human hTERT gene; 2) hTERT
promoter (4kb) region;
6. If foreign gene product(s) will be purified, indicate which foreign gene product will be
purified and briefly describe the procedures for purification, including volumes of culture
N/A
IBC Protocol Form- Jan/2010
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7. If replication-incompetent vectors will be used, include information about how
incompetent vectors are tested for reversion mutations (e.g., endpoint dilution analysis,
plaque assay).
N/A
IBC Protocol Form- Jan/2010
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SECTION C
Hazardous Biological Research Questionnaire
Check 'YES' or 'NO' to the following questions. For any items checked 'YES', include a
thorough description of the work in the following page (question 1).
1. Is agent/material a potential human, animal or plant pathogen?
Yes
Human
Animal
Plant
N/A
2. Is agent/material a toxin?
 If “YES” name of the toxin (s):
 Toxic to:
LD50 :
3. Do you work with quantities (i.e. cultures) larger than 1 liter?
If so, what is the largest volume (liters)?
4. Do you inactivate the agent/material prior to laboratory manipulation?
If “yes”, by what method?
Heat
Chemical
Radiation
Other
Yes
No
No
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
9. Does your work involve a SELECT AGENT? (see appendix H)
If “YES” name the organism (s):
Yes
No
10. Do you work with human blood, tissues, or body fluids?
If “yes” contact Tulane OEHS for guidelines on working with blood-borne
pathogens
Yes
No
5. Do you concentrate the agent/material? If Yes, specify method:
Centrifugation
Precipitation
Filtration
Other
6. Do you expose live animals or humans to the agent/material? If Yes,
specify the Species:
IACUC Approval #
IRB Approval:#
7. Does your project include experiments with Risk Group 2, Risk Group 3 or,
Risk Group 4 agents? *
If yes, which Risk Group?
RG-2
RG-3
RG-4
8. Does your experiment involve work with arthropods (i.e. insects, spiders.
others)? If yes, provide specific details in the next page (question 1)
Work with certain arthropods must conform to the guidelines set forth in the BMBL and
those established by the American Committee of Medical Entomology
11. What is the recommended Biosafety Level for this agent/material? *
BSL-1
BSL-2
BSL-3
BSL-4
*To identify the risk group (RG) classification of the recombinant or etiologic agent(s) and the
proposed biosafety level (See the NIH Guidelines at
http://oba.od.nih.gov/rdna/nih_guidelines_oba.html
And the CDC/NIH Biosafety in Microbiological and Biomedical Laboratories at:
http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm
IBC Protocol Form- Jan/2010
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SECTION C
Hazardous Biological Research Questions
1. Address any item checked ‘YES’ in the Hazardous Biological Research Questionnaire
(include the question number).
2. Describe the key features of the agent/microorganism/toxin or material that you will use
in this project, particularly as it refers to biosafety considerations. Briefly describe the
pathogenesis of the disease caused by this agent.
3. What is the source or your biohazardous agent? (Be specific: i.e. ATCC, CDC, clinical
isolate, etc).
4. Do you work with microorganisms of unknown identity (such as those obtained from soil
or other environmental samples).
5. Does import of this agent into Tulane facilities require USDA/CDC import/transport
permit?
6. Will the agent be cultured/propagated at Tulane? If yes provide a synopsis of the
procedures.
7. Will your work be performed in a BSL3 laboratory? If yes detail the procedures to be
done in the BSL3.
8. Describe any specific biosafety considerations for this agent that have not been addressed
in the previous questions.
IBC Protocol Form- Jan/2010
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Section D
Select Agent Questionnaire


No work with Select Agents can be initiated until IBC approval is obtained
IBC protocols involving Select Agents will be subjected to yearly review.
1. Is there a vaccine available and recommended for persons handling this
Select Agent?
If “yes” will personnel working in this project be offered the vaccine?
Expand the answer to this question under question 1 in the next page
Yes
No
Yes
No
2. Do you plan on shipping, or transporting the select agent?
If “Yes” be aware that a Select Agent must be transported under the
conditions described in specific Code of Federal Regulations (CFRs).
Consult the Office of Biosafety for guidelines.
Yes
No
3. If you are working with a toxin, indicate the largest amount that you will
have in your possession at any given time.
mgs
4. What is the largest volume of culture (bacterial/viral) and approximate
concentration that you will have at any given time? (i.e. 1 liter at 10x106
CFU/ml).
5. Will access to the Select Agent be meticulously controlled at all times?
Yes
No
6. Have you submitted an agent-specific SOP with this application? (This is a
requirement for approval of your protocol).
Yes
No
7. Does your SOP include specific instructions to handle accidental spills and
accidental personnel exposures?
Yes
No
Yes
No
9. Will a copy of the training certification and assessment evaluation for each
person working with the agent be filed with the Biosafety Office?
Yes
No
10. Are you and all the personnel involved in this project proficient in the
general BSL3 SOPs written by the Biosafety Office?
Yes
No
8. Will personnel involved in this project receive agent-specific training, to
make them aware of the risk and hazards associated with this agent, and the
SOPs for accidental spills or exposures?
Who will provide the agent-specific training?
IBC Protocol Form- Jan/2010
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Section D:
Select Agent Questions
1. Provide relevant and detailed information to expand the answers to the above
questionnaire (provide item number)
2. What General SOPs (as provided by the Tulane Biosafety Office) will be used in the
course of these experiments? See Appendix 2 and check all that apply
3. If working with animals indicate the final fate of all animals exposed to the select agent
IBC Protocol Form- Jan/2010
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SECTION E
Biohazards, Decontamination and Training
1. Identify all known and potential hazards or risks associated with the specific
biohazardous agent or with the use of recombinant DNA materials described in this
application (i.e. generation of aerosols, sharps punctures, animal bites, and scratches, etc.)
Biosafety risks in this project (although minimal) are as follows:
- Risk of wounds/punctures with contaminated sharps,
- Contact with human cancer cells,
- Contact with animals tissues. blood or animal waste.
-
2. Describe the necessary facilities/equipment/practices that will be used for all aspects of
the work (for the indicated safety/containment level). (i.e. use of biosafety cabinets ,
aerosol proof centrifuge, aerosolization chambers, etc)
Cell cultures will be done in biosafty cabinets;
Animal works will be done in the vivarium in biosafety cabinets;
BSL2 guidelines will be strictly followed
3. Indicate the personal protective equipment or PPE (gloves, mask, lab coat, PAPR, etc)
required for personnel working in this project.
Personnel working with this project will wear lab coats, gloves for routine work.
4. Specifically describe the decontamination practices to be used. (Lysol, bleach, autoclave,
etc.). Provide detailed description of decontamination for materials/samples that will be
transported out of the laboratory.
All surfaces will be decontaminated with 70% alcohol before and after all procedures
Glassware will be autoclaved
Disposable consumables will be placed in properly labelled bioharzard box
Sharps will be placed in a "sharp containers"
5. Describe the disposal practices of contaminated material, contaminated tissue or animal
carcasses. Include practices for disposal of liquid and solid waste, including sharps.
Disposable waste will be placed in properly labelled bioharzard box
Sharps will be placed in a "sharp containers"
6. For Risk Group 3 agents and for agents that require BSL3 containment, briefly describe
your prior training or experience in this type of work.
N/A
7. Describe how personnel have been (or will be) trained in the handling of the biological
agents described in this application. Who is in charge of training? (The PI is responsible
for keeping accurate records of personnel training)
IBC Protocol Form- Jan/2010
Page 11 of 14
The PI (Dr. Yan Dong) will train all personnel working in this project. Personel will be
instructed on the risks of working with animals, human cancer cells and with recombinant
DNA materials. Personnel will be trained to follow BSL2 guidelines and wear the required
PPE at all times while doing experiments.
8.
If your experiments involve animals, do animal handlers need to be aware of specific
precautions or need to implement changes to the routine animal care?
No changes to routine veterinary care are necessary.
IBC Protocol Form- Jan/2010
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SECTION F:
Investigator’s Assurance
1. I confirm that all persons conducting this work at Tulane University (including students,
fellows, technicians, and collaborators) have been adequately trained in good microbiological
techniques; have received instruction on the specific hazards associated with the work and are
aware of the specific safety equipment, practices, and behaviors required during the course of
the work and use of these facilities.
2. I will report to the Biological Safety Officer immediately any spill of biohazardous material,
any equipment or facility failure (e.g., ventilation failure), and/or any breakdown in
procedure that could result in potential exposure of laboratory personnel and/or the public to
biohazardous material.
3. I confirm that any proposed changes to my work that would result in an increased level of
biohazard will be reported to the IBC before the change is implemented.
4. I confirm that no work requiring IBC approval will be initiated or modified until approval is
received.
5. I have read and understand my responsibilities as Principal Investigator outlined in Section
IV-B-7 of the NIH Guidelines and the Tulane University Institutional Biosafety Committee
(IBC) Policy Manual for the Use of Recombinant DNA, and agree to comply with these
responsibilities.
6. I certify that the information provided within this application is accurate to the best of my
knowledge. I also understand that, should I use the project described in this application as a
basis for a funding proposal (either intramural or extramural), it is my responsibility to
ensure that the description of the work in the funding proposal is identical in principle to that
contained in this application.
Yan Dong
Typed Name of Principal Investigator
Signature of Principal Investigator
Date
UPON APPROVAL YOU MUST SEND A SIGNED COPY OF THIS LAST PAGE TO THE IBC
OFFICE (by Campus mail Box TW-5, Fax 8-0370, or email (scanned copy) to ibc@tulane.edu)
For IBC Use Only
Protocol Number:
Protocol Title:
Experiment Class Determination (check one)
III-A Requires IBC approval, RAC review, and NIH Director approval before initiation
III-B Requires NIH/OBA and IBC approval before initiation
III-C Requires IBC and IRB approvals and RAC review before research participant enrollment
III-D Requires IBC approval before initiation
III-E Requires IBC notice simultaneous with initiation
III-F Exempt
IBC Protocol Form- Jan/2010
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IBC Signature:
Date:
IBC Protocol Form- Jan/2010
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