Periodic Reporting for period 1 - MAGNOSTICS (Biomimetic MAGnetic nanoparticles with homotypic targeting for combinatorial theraNOSTICS of triple-negative breast cancers)
Reporting period: 2023-05-02 to 2025-05-01
MAGNOSTICS aimed at developing biomimetic platform for the targeted delivery of siRNA against ABCB1 gene and anti-cancer drug to achieve a multi-modal thernaostics for triple negative breast cancer (TNBC). Metastatic TNBCs are aggressive tumors with poor prognosis contributing to about 15 % of all the breast cancers with high mortality rates. Characterized by the absence of estrogen, progesterone, and human epidermal growth factor 2 receptors, endocrine and targeted therapies are ineffective. Thus, chemotherapy is still the cornerstone for TNBC treatment. However, chemotherapy is strongly impeded by non-specificity and efflux/drug resistance, which lead to recurrence and relapse. In metastatic cases, 90% of therapy failure is attributed to chemoresistance. ATP-binding cassette (ABC) transporters are one of the strong reasons leading to drug efflux and thus chemoresistance. Suppressing the expression of ABCB1 gene with a small-interfering RNA (siRNA) can inhibit drug efflux. A combined treatment approach addressing drug efflux with superior synergistic effect is required for the improved management of TNBCs. Through MAGNOSTICS, we aimed to develop multimodal biomimetic nanoplatform to address non-specificity and ABCB1-mediated chemoresistance in TNBC. Thus, the objectives of MAGNOSTICS were to synthesize and characterize biomimetic magnetic iron-oxide nanoparticles (MNPs) loaded with the anticancer drug, doxorubicin (Dox) and siRNA against ABCB1 gene with a coating of cancer cell membrane (CM) obtained from a drug-resistant TNBC cell line. Through MAGNOSTICS, the biomimetic nanoparticles (NPs) were found to be biocompatible, demonstrated efficient uptake by cancer cells, and displayed immune escape capability. Importantly, these biomimetic NPs showed excellent gene silencing and retention of dox. Furthermore, with the magnetic hyperthermia, the NPs demonstrated enhanced siRNA release and as MRI contrast agents.
To achieve the overall goal of MAGNOSTICS, the following objectives were focused;
1. To synthesize and characterize CM-coated MNPs conjugated with CDDP and siABCB1. The iron-oxide NPs were loaded with siRNA and further intercalated with Dox and finally coated with cell membrane. The biomimetic NPs were successfully characterized for size, surface charge, morphology and conjugation by DLS, TEM, and FT-IR. The coating was also confirmed by determining the presence of specific proteins on the CM coated NPs.
2. To study the biocompatibility/uptake and single therapeutic strategies as a proof-of-concept in 2D cell culture. In this objective, the cell membrane coated NPs were studied for their stability, uptake, and single therapeutics. The NPs showed excellent compatibility to the non-cancerous cells and demonstrated good uptake and potential for multi-modal approach for theranostic.
3. To develop 3D multicellular tumor spheroids and demonstrate the effectiveness of biomimetic nanoparticles as a combined theranostic platform. The ability of NPs to silence the gene, retain the drug and enhance the cancer cell death by magnetic hyperthermia on spheroids is being tested.
4. To ensure further professional development of the ER through training, dissemination and public engagement. Through this objective, the goal and objectives of MAGNOSTICS were disseminated to the scientific community via different conferences and communicated to public at open days, events, and interviews.
1. To synthesize and characterize CM-coated MNPs conjugated with CDDP and siABCB1. The iron-oxide NPs were loaded with siRNA and further intercalated with Dox and finally coated with cell membrane. The biomimetic NPs were successfully characterized for size, surface charge, morphology and conjugation by DLS, TEM, and FT-IR. The coating was also confirmed by determining the presence of specific proteins on the CM coated NPs.
2. To study the biocompatibility/uptake and single therapeutic strategies as a proof-of-concept in 2D cell culture. In this objective, the cell membrane coated NPs were studied for their stability, uptake, and single therapeutics. The NPs showed excellent compatibility to the non-cancerous cells and demonstrated good uptake and potential for multi-modal approach for theranostic.
3. To develop 3D multicellular tumor spheroids and demonstrate the effectiveness of biomimetic nanoparticles as a combined theranostic platform. The ability of NPs to silence the gene, retain the drug and enhance the cancer cell death by magnetic hyperthermia on spheroids is being tested.
4. To ensure further professional development of the ER through training, dissemination and public engagement. Through this objective, the goal and objectives of MAGNOSTICS were disseminated to the scientific community via different conferences and communicated to public at open days, events, and interviews.
During the lifetime of the project, activities related to addressing the aim and related objectives were carried out. The first and an important objective was to synthesize and characterize cell membrane cloaked nanoparticles loaded with anti-cancer drug and siRNA. Different methods were tried for the synthesis of iron-oxide nanoparticles, with the hydrothermal method finalized. The DOX showed excellent intercalation in siRNA with high loading efficiency. The cell membrane coating onto the NPs was tried using extrusion and sonication method. The pattern of the proteins in CM corroborated with that from the cell membrane coated NPs. The nanoparticles showed efficient loading of siRNA and DOX. Release studies demonstrated enhanced release of siRNA without and with magnetic hyperthermia, confirming the effect of hyperthermia on the release profile. The NPs were found to be internalized by the cancer cells, which demonstrated homotypic targeting of these nanoparticles. The nanoparticles also showed biocompatibility towards healthy breast cancer cells, with immune escape properties owing to the coating with cell membrane. With regards to gene therapy, the nanoparticles have shown optimum ABCB1 silencing with retention of DOX. This multimodal approach of MAGNOSTICS wherein the hyperthermia, gene silencing and chemotherapy provide a synergistic effect to control the cancer growth has a potential future for other cancer types and different health conditions.