M.Sc Thesis


M.Sc StudentHarris Yuval
SubjectNanoparticles for KRAS Driven Lung Cancer-Development,
Optimization and Automation
DepartmentDepartment of Biomedical Engineering
Supervisor DR. Yosef Shamay


Abstract

Lung cancer is the most common type of cancer, and one of the leading causes of death globally, with a 5-years-surviavel rate of only 22%. There are two main types of lung cancer- Small Cell Lung Cancer (SCLC) and Non-SCLC (NSCLC) which is the focus of this thesis as it represents 85% of the cases. The current treatments for NSCLC include radiation, surgical intervention, immunotherapy, chemotherapeutic drugs, and personalized kinase inhibitors, depending on the stage, protein and genetic markers. Even though for some genetic drivers of NSCLC, such as ALK and EGFR, a personalized approach is clinically beneficial, for the most frequently mutated oncogene, KRAS, there is no such therapy. KRAS is also frequently mutated in many other cancers, including pancreatic and colon cancer. One out of four cases of lung cancer harbor a mutation in the KRAS gene and is commonly associated with poor prognosis.

As mutated KRAS leads to constant activation of multiple kinases signaling cascades, such as MAP kinase and PI3 kinase, it was hypothesized that kinase inhibitors (KIs) might have clinical benefits. KIs are small molecule, hydrophobic drugs which are limited to systemic oral administration and result in dose limiting side-effects and acquired drug resistance which might explain the lack of clinical efficacy.

It was suggested that by trapping KIs in nanoparticles (NPs), the drug can be delivered selectively to the tumors via the enhanced permeability and retention (EPR) effect and reduce toxicity and avoid drug resistance. A recent study has showed that a subset of sulfated indocyanine dyes self-assemble with hydrophobic drugs to form stable NPs. These novel NPs have high drug loading which may further advance the effectiveness of NPs in cancer treatments, especially for KIs. Nevertheless, the process of preparing and purifying multiple NPs with different drugs and dyes in a manual way, for optimal formulation is challenging and time consuming.

In the study presented here, we developed an automated procedure to prepare and purify NPs-based dye-drug combinations with high drug loading. We discovered a novel dye stabilizer, named IR595 which is highly effective for KIs relevant for the KRAS pathway and tested this novel NP system in vitro and in vivo mouse model of lung cancer. In addition, we developed a KRAS driven, drug resistant cell line and found two novel drug combinations, which can self-assemble to drug-dye NPs, that avoid acquired resistance. All of our data supports the hypothesis that our new NPs is safe and effective in vivo for personalized lung cancer-based nanomedicine.