Gel NPs dispersed in DI water indicated an average hydrodynamic diameter of 225??20?nm having a polydispersity index (PDI) less than or equal to 0.1. In the presence of TKI, GAB1-SHP2 dissociation happens, leading to cell death. The outcome of this study provides a encouraging platform for treating NSCLC individuals harboring KRAS mutation. NSCLC is definitely diagnosed in an estimated 220,000 individuals each year with five-year overall survival rates of 16 percent1. A recent statement confirmed that 16 percent of NSCLC individuals carry oncogenic KRAS mutation2. A potent drug targeted against KRAS mutation has not yet been developed and the objective response rate with the current standard of care is just three percent. An earlier report had suggested siRNA therapy renders the undruggable KRAS mutant cells to become susceptible to Tyrosine Kinase Inhibitors (TKI)3. Short interfering RNA (siRNA) is definitely a well-known approach for effecting gene therapy to provide subsequent sensitization towards complementary restorative agents. However, stable delivery of siRNA is definitely a significant challenge due to its high degradation rate in the presence of serum proteins and enzymes. To conquer this challenge, several nanoparticle centered carrier systems have been attempted and those include retroviral vectors, 7ACC1 liposomes, polymeric, and metallic nanoparticles3,4,5,6,7. In these reported studies the physicochemical and surface properties of the particle were modified for delivering the siRNA to cytoplasm of the infected cells. Regrettably, these nanoparticles suffer from serious limitations such as stability issues during synthesis, premature launch in serum, inefficient endosomal escape, and interferon response4,8,9. Importantly, oncogene knockdown only has less impact on the malignancy cell apoptosis since the cells tend to adopt another effector pathway for survival3,10,11,12. Consequently, a need for complementary drug for initiating the apoptosis post knockdown is needed. Indeed, drugging cells separately and exogenously post oncogene knockdown has been reported earlier9,10,11. A combined delivery system wherein, co-delivery of a drug along with siRNA to impede growth and survival of the cell has also been attempted13. The relevance of the combined delivery is to ensure the complementary drug enters the same cells that are affected by siRNA at a predetermined appropriate proportion and time for causing cellular apoptosis. However, incorporation of siRNA (with minimal degradation) having a drug and a biomarker-targeting antibody into a solitary platform is definitely synthetically challenging. Therefore, stable and targeted delivery with concomitant cytotoxic action to malignancy cells continues to be at early exploratory phases. Significant efforts have been made to Rabbit Polyclonal to CPA5 understand the downstream effect of oncogene knockdown mediated via siRNA till day14. Malignancy cells have several parallel operating pathways, with one main effector pathway coupled to several parallel effector pathways15. The parallel pathways remain dormant until the working pathway is definitely disrupted. Switch in the protein expression levels upon knock down of oncogene present in the primary pathway results in switch of downstream protein and gene manifestation levels controlled by complex cellular mechanism. This mode of intra-cellular functioning adaptation evolves to drug resistance within malignancy cells that are previously responding to therapy16. On the other hand, KRAS mutant adenocarcinoma of NSCLC have been undruggable till day17. While mutations happen at variation position of KRAS, oncogenic effect at codon 12 (Glycine-12 to Cysteine, G12C) of KRAS is the most commonly happening mutation and yet to receive a dedicated drug18. Although, in recent times, few attempts have been made for focusing on G12C mutation through a small molecule inhibitor, RNAi therapy is definitely emerging like a encouraging tool that may be applied across all types of mutations supplemented with currently approved medicines19,20. For example, knocking down a specific gene of undruggable malignancy, such as KRAS mutant adenocarcinoma of NSCLC, can activate a parallel dormant effector pathway that may be sensitive to a TKI3. In this work, we statement the synthesis and utilization of gelatin nanoparticle (Gel NP) like a carrier system encapsulated with gefitinib (GelGEFNP). The Gel NP is definitely surface functionalized with cetuximab (Ab), a EGFR focusing on antibody, conjugated to KRAS G12C specific siRNA (Ab-siRNA conjugate) (Fig. 1). KRAS G12C specific siRNA is definitely chemically conjugated to cetuximab by thio-ether linkage. Cetuximab antibody target EGFR receptor within the cells and also protects siRNA from external degradation. We believe the 14?kDa siRNA is camouflaged by 146?kDa cetuximab antibody thus remaining un-exposed to external environment. The knockdown 7ACC1 of 7ACC1 oncogene upon treatment with TBN was confirmed by studying downstream signaling.