Dual targeting of VEGFA and PDGFb in cervical cancer: a possible antiangiogenic therapeutic tool using receptor specific CPPs

Title
Dual targeting of VEGFA and PDGFb in cervical cancer: a possible antiangiogenic therapeutic tool using receptor specific CPPs

Funding: UEFISCDI
Contract No. : 102/12.01.2012


Project summary
Cervical cancer is one of the most aggressive gynecological diseases, mainly due to rapid development of a functional tumor vascular network, a process termed “angiogenesis”. Angiogenesis is maintained by a balance of pro and antiangiogenic factors, that in cancer are inclined towards proangiogenesis. VEGFA and PDGFb are two of the most important proangiogenic factors, and are upregulated in cervical cancer. Although tremendous efforts have been put in this field, mono-antiangiogenic therapy has failed, which led to the ideea of multiple targeting. This project is focused on using RNA interference technology as a possible therapeutic tool in cervical cancer. Using RNA interference as therapeutic tool has mostly been limited by the lack of effective delivery systems. Cell penetrating peptides have been proved to be able to translocate across cell membrane different types of cargoes, including siRNA, without any associated cytotoxicity. We propose an interdisciplinary approach of the two main problems in cancer therapy: the need for effective antiangiogenic agents and a targeted delivery system that would ensure the transport of these agents to and into tumors without affecting healthy cells. Moreover, the proposed molecular studies will shade new light in the molecular and cellular mechanism of combined effects of VEGF and PDGF inhibition in cervical tumor cells.

Objectives
General objective: using RNA interference technology as a possible therapeutic tool in dual targeted antiangiogenic therapy in cervical cancer

Principal objectives

Objective 1. Evaluation of the cellular effect of dual targeting of VEGFA and PDGFb as a possible antiangiogenic strategy in cervical cancer cells.
Objective 2. Evaluation of the molecular mechanism of dual targeting of VEGFA and PDGFb as a possible antiangiogenic strategy in cervical cancer cells.
Objective 3. Rational design and testing of a tumor targeted delivery system for siRNA transport into VEGF receptor expressing tumor cells.

Budget 300000 RON

Work plan:
Objective 1: Experimental design and setup
Activity 1.1. Treatment optimization

Objective 2: Evaluation of cellular effects of siRNA treatment
Activity 2.1. Apoptosis and angiogenesis cell assays
Activity 2.2. Data integration and analysis

Objective 3: Uptake evaluation of the designed CPPs (4-15)
Activity 3.1. Uptake evaluation of VEGFR+ cells
Activity 3.2. Uptake evaluation of VEGFR-cells

Objective 4: Evaluation of molecular effects of siRNA treatment
Activity 4.1. PCR array analysis
Activity 4.2. Protein quantification
Activity 4.3. Data integration and analysis

Objective 5: Evaluation of the antiangiogenic effects induced by the designed CPPs
Activity 5.1. Apoptosis and angiogenesis cell assays
Activity 5.2. Data integration and analysis

Objective 6: Result dissemination
Activity 6.1. Attend three conference, publish two ISI article

Objective 7: Project management
Activity 7.1. Monitoring, reports preparation

Preliminary results:

1. Tested CPPs are more efficient and less toxic in siRNA delivery compared to some commercial agents, maximum effect is at 48 hours post treatment
2. Both VEGFA and PDGFB silencing induce several biological effects, a synergistic effect is observed when dual targeting is employed
3. Targeted CPPs are efficient in siRNA delivery both in VEGFRs+ and VEGFR- cells.


Dissemination:
Poster presentation: O. Tudoran, et al., Transcriptional mechanism of apoptosis induced by VEGFA inhibition in cervical cancer cells, The Tumor Microenvironment, Dublin, 17-19 September 2012



Project director: Oana Tudoran, Phd
Tel 0264590638
E-mail: oana.tudoran@iocn.ro