'Microvesicle-mediated intercellular communication involving cancer stem cells.' Undergraduate Research Project Application Form.

INSTRUCTIONS - PROFESSORS: Fill out Sections A & B then submit this form online. (You will receive an email copy of the form. The Office for Undergraduate Research in Science will also post the project online, indicating whether the project is open for students to apply or taken.) DONE

INSTRUCTIONS - STUDENTS: You may receive this form by email, or you may download it after it has been posted. Either way, print this form. Complete and sign Section C on the hardcopy. Ask your supervisor to sign Section D. Take it to the department corresponding to the course number in Section A (this may or may not be your own department). Do not register for a '396' course on Minerva until you receive departmental permission. Have a discussion with your supervisor about time/work expectations, keeping in mind that this is a 3-credit course (roughly, 10 hours per week for 12 weeks). Remember that a '396' course is an elective.

INSTRUCTIONS - DEPARTMENTS: After the unit chair/director/designate approves (or not) this project, notify student. If approved, please give student permission to register on Minerva, and fax this form (with signatures) to the Office for Undergraduate Research in Science.

QUESTIONS OR FEEDBACK? Contact Victor Chisholm by email, or phone 514-398-5964.

SECTION A: SUPERVISOR INFORMATION

Name: Janusz RAK

Email: janusz.rak [at] mcgill.ca (janusz dot rak at mcgill dot ca)

Phone: 514-412-4400 x 22342

Website:

Supervisor's department: Biochemistry

Supervisor's department (if none of the above)

Course number: BIOC396 (Biochemistry)

SECTION B: PROJECT INFORMATION

Term: Winter 2010-2011

Project start date: January 4th, 2011

Project end date: April 8th, 2011

Project title: Microvesicle-mediated intercellular communication involving cancer stem cells

Project description: Cancer cells communicate with each other and their surrounding (and distant) normal counterparts by releasing membrane organelles called microvesicles (MVs). MVs contain a wealth of bioactive cargo, including oncoproteins, their RNA transcripts and oncogenic microRNA, all of which could be released and transferred to other cells. Such molecular transfer may trigger proliferation, migration proangiogenic activity and reprogramming of cellular properties, thereby synchronizing multicellular behaviour.

Aggressive brain tumours known as glioblatoma multiforme (GBM) are composed of heterogeneous cellular populations that originate from, and are maintained by a small subset of cells known as tumour initiating cells (BTICs), or cancer stem cells (CSCs). CSCs communicate with their surroundings, but whether this involves MVs remains unknown. MV production by BTICs is being studied in the Rak laboratory using a murine BTIC cell line (mBTIC), which harbours unique genetic alterations (Ink4A-/-/EGFRvIII) and is able to give rise (differentiate) to proliferative non-CSC progeny.

I hypothesize that MV production, content and biological activity changes as BTICs transit to their proliferative descendants devoid of the tumour initiating potential. I will contribute to addressing these questions by realizing 2 experimental aims:

Aim 1. I will transfect mBTIC cells with cDNA encoding myristoylated green fluorescent protein (myr-GFP). I will test under fluorescent microscope and by flow cytometry the expression of GFP by these cells and production of GFP-containing MVs.

Aim 2. I will measure how the release of MVs changes when mBTICs differentiate to their progeny in vitro. The cells will be switched from the stem cell media, in which mBTICs are maintained as neurospheres, to serum containing monolayer cultures where they become more differentiated non-CSCs. I will collect conditioned medium from both cultures and measure MV-associated fluorescence using the Nanosight particle counter under various conditions. I will also test the content of EGFRvIII oncogene and other markers in these various MVs (Western). The future plan (likely beyond my tenure) is to extend these studies to in vivo conditions where mBTIC are implanted to mice and grow as tumours with spontaneous generation of non_CSCs. Both cell types are expected to emit MVs into their surrounding tissue, cause the uptake of GFP by normal cells, and emerge as circulating MVs in blood, from which they can be isolated and analysed.

Significance. My project will help create a simple model, with which to study vesiculation of cancer stem cells.

Prerequisite: 1 term completed at McGill + CGPA of 3.0 or higher; or permission of instructor.

Other prerequisite, if applicable:

Grading scheme (The final report must be worth at least 50% of final grade): Final grade shall be based on laboratory performance as evaluated by the research supervisor (50%) and the final written research report (minimum 10 pages) graded by the supervisor and the course coordinator or the coordinator's delegate (50%).

Other project information: This project involves cell culture and some basic biochemical techniques (Western)

Project status - This project is: Taken. The professor has no more '396' projects this term.

How students can apply: N/A; this project is filled.

If other, please specify:

Ethics, safety, and training

Which of the following, if any, is involved? One or more of the following

Animal subjects [ ]

Human subjects [ ]

Biohazardous substances [x]

Radioactive materials [ ]

Handling chemicals [x]

Using lasers [x]

Supervisors are responsible for the ethics and safety compliance of undergraduate students.

SECTION C: STUDENT INFORMATION

Do not complete this section unless/until the student is identified.

Name:

McGill ID:

Email (first.last [at] mail.mcgill.ca):

Phone:

Program (E.g., B.Sc. Maj. Chem. Min. Biol.):

Level (U0 / U1 / U2 / U3):

Student signature - I have not applied for another '396' course in this term:

SECTION D: APPROVALS.

Do not complete this section unless/until the student is identified.

Supervisor: I give my permission for the student identified in section C to register for this project under my supervision.

Supervisor's signature:

Date:

Unit chair/director/designate: I certify that this project conforms to departmental requirements for 396 courses.

Unit chair/director/designate's name:

Unit chair/director/designate's signature:

Date: