Chapter 6
Magnetic Nanoparticles as a Carrier of dsRNA for Gene
Therapy
Małgorzata Grabowska, Bartosz F. Grzes ´ kowiak, Katarzyna Rolle,
and Radosław Mro ´ wczyn ´ ski
Abstract
Glioma belongs to the most aggressive and lethal types of cancer. Glioblastoma multiforme (GBM), the
most common type of malignant gliomas, is characterized by a poor prognosis and remains practically
incurable despite aggressive treatment such as surgery, radiotherapy, and chemotherapy. Brain tumor cells
overexpress a number of proteins that play a crucial role in tumorigenesis and may be exploited as
therapeutic targets. One such target can be an extracellular matrix glycoprotein—tenascin-C (TN-C).
Downregulation of TN-C by RNA interference (RNAi) is a very promising strategy in cancer therapy.
However, the successful delivery of naked double-stranded RNA (dsRNA) complementary to TN-C
sequence (ATN-RNA) requires application of delivery vehicles that can efficiently overcome rapid degradation by nucleases and poor intracellular uptake. Here, we present a protocol for application of MNP@PEI as
a carrier for ATN-RNA to GBM cells. The obtained complexes consisted of polyethyleneimine (PEI)coated magnetic nanoparticles combined with the dsRNA show high efficiency in ATN-RNA delivery,
resulting not only in significant TN-C expression level downregulation, but also impairing the tumor cells
migration.
Key words Magnetic nanoparticles, RNAi, Tenascin-C, Glioblastoma, Gene therapy, Cytotoxicity,
Transfection, Double-stranded RNA
1 Introduction
The most common type of glioma, accounting for 55% of brain
tumors, is glioblastoma multiforme (GBM). It is rated as the most
aggressive, a fourth grade cancer, according to the World Health
Organization classification [1]. Recommended therapies include
surgical resection, radiotherapy, and systemic chemotherapy with
oral prodrug temozolomide. Depending on the therapy, a median
survival rate amounts 12–15 months, while failure to attempt treatment results in patient’s death within 3 months [2].
None of the current state-of-the-art treatments for malignant
glioma can be regarded as effective. Complete surgical resection of
Kumaran Narayanan (ed.), Bio-Carrier Vectors: Methods and Protocols, Methods in Molecular Biology, vol. 2211,
https://doi.org/10.1007/978-1-0716-0943-9_6, © Springer Science+Business Media, LLC, part of Springer Nature 2021
69
Magnetic Nanoparticles as a Carrier of dsRNA for Gene
Therapy
Małgorzata Grabowska, Bartosz F. Grzes ´ kowiak, Katarzyna Rolle,
and Radosław Mro ´ wczyn ´ ski
Abstract
Glioma belongs to the most aggressive and lethal types of cancer. Glioblastoma multiforme (GBM), the
most common type of malignant gliomas, is characterized by a poor prognosis and remains practically
incurable despite aggressive treatment such as surgery, radiotherapy, and chemotherapy. Brain tumor cells
overexpress a number of proteins that play a crucial role in tumorigenesis and may be exploited as
therapeutic targets. One such target can be an extracellular matrix glycoprotein—tenascin-C (TN-C).
Downregulation of TN-C by RNA interference (RNAi) is a very promising strategy in cancer therapy.
However, the successful delivery of naked double-stranded RNA (dsRNA) complementary to TN-C
sequence (ATN-RNA) requires application of delivery vehicles that can efficiently overcome rapid degradation by nucleases and poor intracellular uptake. Here, we present a protocol for application of MNP@PEI as
a carrier for ATN-RNA to GBM cells. The obtained complexes consisted of polyethyleneimine (PEI)coated magnetic nanoparticles combined with the dsRNA show high efficiency in ATN-RNA delivery,
resulting not only in significant TN-C expression level downregulation, but also impairing the tumor cells
migration.
Key words Magnetic nanoparticles, RNAi, Tenascin-C, Glioblastoma, Gene therapy, Cytotoxicity,
Transfection, Double-stranded RNA
1 Introduction
The most common type of glioma, accounting for 55% of brain
tumors, is glioblastoma multiforme (GBM). It is rated as the most
aggressive, a fourth grade cancer, according to the World Health
Organization classification [1]. Recommended therapies include
surgical resection, radiotherapy, and systemic chemotherapy with
oral prodrug temozolomide. Depending on the therapy, a median
survival rate amounts 12–15 months, while failure to attempt treatment results in patient’s death within 3 months [2].
None of the current state-of-the-art treatments for malignant
glioma can be regarded as effective. Complete surgical resection of
Kumaran Narayanan (ed.), Bio-Carrier Vectors: Methods and Protocols, Methods in Molecular Biology, vol. 2211,
https://doi.org/10.1007/978-1-0716-0943-9_6, © Springer Science+Business Media, LLC, part of Springer Nature 2021
69