'Assessment of the role of mTor signaling in fludarabine resistance in CLL' - ANAT 396 Undergraduate Research Project Application Form.

INSTRUCTIONS - PROFESSORS: Please review this form. Complete or correct the sections, as applicable, from "Supervisor's Name" to "Ethics, safety, and training". Please sign and date near the bottom ("Supervisor's signature").

INSTRUCTIONS - STUDENTS: You may receive this form by email, or you may download it fter it has been posted. Either way, print and review this form. Complete or correct the sections, from "Student's Name" to "Student's Level", and sign ("Student signature). Ask your supervisor to sign her/his section near the bottom. Take it to the department corresponding to the course number; 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, please 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. The Office for Undergraduate Research in Science will later post the project online, indicating whether the project is open for students to apply or taken.

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


Supervisor's Name: Raquel Aloyz

Supervisor's Email: raquel.aloyz [at] mcgill.ca

Supervisor's Phone: 514-340-8222 ext. 5288

Supervisor's Website: https://mcgill.ca/translational-research-cancer/researcher-biographies/aloyz and http://www.ladydavis.ca/en/raquelaloyz

Supervisor's department: Department of Oncology

Course number: ANAT396 (Anatomy and Cell Biology)

Term: Fall 2011-2012

Project start date: Wednesday, September 1, 2011

Project end date: Tuesday, December 6, 2011

Project title: Assessment of the role of  mTor signaling in fludarabine resistance in CLL

Project description: Targeting eIF4E signaling in primary CLL lymphocytes

Chronic lymphocytic leukemia (CLL), the most common leukemia in adults in the western world, is an incurable disease.  Although CLL patients might not require treatment for years, one third of patients with aggressive disease require early intervention upon diagnosis. In addition, even though the patients initially respond to available treatments, they develop resistance and relapse. [1]. The mammalian target of rapamycin (mTOR), a downstream effector of AKT, has been identified as a drugable target in hematological malignancies, including CLL [2] [3]. Nevertheless, mTor inhibitors failed to induce significant apoptosis of either cycling or quiescent cells and showed so far modest clinical responses associated with toxicity [4] [2]. This may be due to their ability to block the feedback activation of PI-3k resulting from mTorc-1 inhibition [1]. Another approach is to target mTorc-1 downstream targets such as the translation factor eIF4E [5]. Ribavirin is a well-characterized nucleoside analogue with broad antiviral activity that has also been shown to target eIF4E in a variety of systems, thereby inhibiting translation and/or mRNA export of eIF4E dependent transcripts [6-8]. We propose to assess the effect of ribavirin on the survival of primary CLL lymphocytes isolated from the peripheral blood of affected patients diagnosed at the Hematology Clinic (Jewish General Hospital) after consent. We will utilize the maximal clinically achievable dose of ribavirin (10M) alone or in combination with fludarabine (FLU). FLU, the first line treatment for the disease, is a purine analog that has been shown to induce DNA damage in CLL lymphocytes. FLU resistance has been associated with an altered balance in the expression of antiapoptotic and proapoptotic factors such as bcl2, MCl-1 and Bax [9] [10].

Specific Objectives:

  • We will test CLL lymphocytes isolated from the blood of 10-15 patients
  • The lymphocytes will be treated in vitro with FLU alone (0-30uM), ribavirin alone (10uM) of fludarabine in combination with 10uM ribavirin. Cytotoxicity will be assessed using the MTT assay and induction of apoptosis using expression of annexinV and/or active aspase-3 by flow cytometry.
  • The steady state of activation of the mTor pathway (phosphorylated mTor,  AKT and eIF4E) will be assessed by western blot 24 and 48 hours after in vitro treatment with fludrabine IC50 concentration plus vehicle and fludarabine. Expression of know targets downstream eIF4E involved in CLL survival/resistance to chemotherapy will be assessed by western blot as well including bcl2 and Mcl-1.

REFERENCES

1.      Wickremasinghe, R.G., A.G. Prentice, and A.J. Steele, Aberrantly activated anti-apoptotic signalling mechanisms in chronic lymphocytic leukaemia cells: clues to the identification of novel therapeutic targets. Br J Haematol, 2011. 153(5): p. 545-56.

2.      Ringshausen, I., C. Peschel, and T. Decker, Mammalian target of rapamycin (mTOR) inhibition in chronic lymphocytic B-cell leukemia: a new therapeutic option. Leuk Lymphoma, 2005. 46(1): p. 11-9.

3.      Zent, C.S., et al., The treatment of recurrent/refractory chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL) with everolimus results in clinical responses and mobilization of CLL cells into the circulation. Cancer, 2010. 116(9): p. 2201-7.

4.      Decker, T., et al., A pilot trial of the mTOR (mammalian target of rapamycin) inhibitor RAD001 in patients with advanced B-CLL. Ann Hematol, 2009. 88(3): p. 221-7.

5.      Hsieh, A.C., M.L. Truitt, and D. Ruggero, Oncogenic AKTivation of translation as a therapeutic target. Br J Cancer, 2011. 105(3): p. 329-36.

6.      Tan, K., et al., Ribavirin targets eIF4E dependent Akt survival signaling. Biochem Biophys Res Commun, 2008. 375(3): p. 341-5.

7.      Kentsis, A., et al., Ribavirin suppresses eIF4E-mediated oncogenic transformation by physical mimicry of the 7-methyl guanosine mRNA cap. Proc Natl Acad Sci U S A, 2004. 101(52): p. 18105-10.

8.      Assouline, S., et al., Molecular targeting of the oncogene eIF4E in acute myeloid leukemia (AML): a proof-of-principle clinical trial with ribavirin. Blood, 2009. 114(2): p. 257-60.

9.      Kitada, S., et al., Expression of apoptosis-regulating proteins in chronic lymphocytic leukemia: correlations with In vitro and In vivo chemoresponses. Blood, 1998. 91(9): p. 3379-89.

10.     Podhorecka, M., et al., Resveratrol increases rate of apoptosis caused by purine analogues in malignant lymphocytes of chronic lymphocytic leukemia. Ann Hematol, 2011. 90(2): p. 173-83.

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

Grading scheme (The final report must be worth at least 50% of final grade): A final report worth at least 50%. Exact details to be determined.

Other project information:

  1. This project will be co-supervised with Dr. Chantal Autexier. Departments of Anatomy and Cell Biology and Medicine, Division of Experimental Medicine, Telephone: (514) 340-8222, ext. 4651, e-mail address: chantal.autexier [at] mcgill.ca (, )website: http://people.mcgill.ca/chantal.autexier/ and http://www.ladydavis.ca/en/chantalautexier
  2. The student is required to take the biosafety training course offered at the Lady Davis Institute.

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

Ethics, safety, and training: Supervisors are responsible for the ethics and safety compliance of undergraduate students. Which of the following, if any, is involved? Biohazardous substances

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

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