Periodic Reporting for period 1 - PiezoMagBBB (Noninvasive Modulation of the Blood Brain Barrier using PiezoMagnetic Carbon Nanoneedles)
Reporting period: 2020-09-21 to 2022-09-20
There is a significant number of diseases affecting the brain but current treatments for these conditions are limited to therapies which only reduce the symptoms of the disease but don't halt its progression or treat it. This is because it is very hard for medicines to reach the brain due to the presence of a tight barrier known as the blood brain barrier. BBB protects the integrity of the brain by controlling the flux of molecules into the brain parenchyma. This defense barrier however presents a great challenge for getting therapeutic molecules into the brain for the treatment of brain tumours and neurological disorders. Therefore, there is a pressing need for the development of novel drug delivery systems (DDS) to shuttle drugs across the BBB.
The overall aim of PiezoMagBBB was to introduce a new delivery system to cross the Blood Brain Barrier (BBB) by developing a smart nano-construct named PiezoMagnetic Carbon Nanoneedles (PMCNNs) which will act as nanotransducers for converting short-wave ultrasound (US) into electric pulses to electrically permeate the BBB.
As part of the project, an ultrasound responsive, non-invasive, BBB penetrating nanosystems were developed and their therapeutic efficacy as delivery systems for anticancer therapeutics in 3D Glioblastoma Models was evaluated. Thus, the proposed research provides innovative nanocarriers that can deliver a wide range of therapeutics inside the brain for the treatment of neurodegenerative disorders as well as brain cancer.
The overall aim of PiezoMagBBB was to introduce a new delivery system to cross the Blood Brain Barrier (BBB) by developing a smart nano-construct named PiezoMagnetic Carbon Nanoneedles (PMCNNs) which will act as nanotransducers for converting short-wave ultrasound (US) into electric pulses to electrically permeate the BBB.
As part of the project, an ultrasound responsive, non-invasive, BBB penetrating nanosystems were developed and their therapeutic efficacy as delivery systems for anticancer therapeutics in 3D Glioblastoma Models was evaluated. Thus, the proposed research provides innovative nanocarriers that can deliver a wide range of therapeutics inside the brain for the treatment of neurodegenerative disorders as well as brain cancer.
The overall aim of the project has been met by completion of the four research objectives (ROs) and associated work packages (WPs), as described.
The first research objective was to rationally design and characterise novel PMCNNs and this objective was met by the completion of WP1 by the preparation and characterisation of PMCNNs. For that first PiezoMagnetic nanoparticles (PMNPs) were fabricated and attached onto the f-CNTs through electrostatic and covalent functionalisation.
The second research objective was to evaluate the in vitro cytotoxicity of PMCNNs. This objective was met by the completion of WP2 by performing the toxicological evaluation of PMCNNs in different BBB cells Briefly the effect of concentration and incubation time of PMCNNs and their components were studied in vitro in different types of brain cells (human primary astrocytes and human brain microvascular endothelial cells etc.). Here we mainly used the modified lactate dehydrogenase (LDH) assay to circumvent the interference between carbon-based nanoparticles and fluorophores.
The third research objective was to investigate and elucidate the uptake mechanism(s) and fate of PMCNNs in BBB models. To achieve this objective, we successfully completed by assessing the translocation of PMCNNs in BBB models. Briefly a trans-well co-culture BBB model and multicellular spheroidal BBB model were developed to study the translocation of PMCNNs. The tightness of the BBB models was measured by imaging the expression of transmembrane proteins such as Claudin, Occludin and Zonula Occludens along with measuring the trans-endothelial electrical resistance (TEER) of the Transwell BBB models using an EVOM epithelial voltohmmeter. Cellular uptake of PMCNNs (25µg/ml) in BBB co-culture model were investigated in the presence and absence of ultrasound (US). The uptake of PMCNN was tracked through two photon microscopy imaging due to the intrinsic second harmonic generation imaging property of PMCNNs.
The fourth and final research objective was to evaluate the potential of PMCNNs as drug delivery system for the delivery of exemplar anticancer drugs in 3D Glioblastoma model. This objective was met by the completion of WP4 by evaluating the therapeutic potential of PMCNNs as delivery systems for anticancer therapeutics in 3D Glioblastoma Models. Briefly we assessed the therapeutic effects of PMCNNs as delivery systems for the anticancer drug cisplatin (CDDP) in 3D glioblastoma tumour spheroid models. Cisplatin (CDDP) was loaded onto PMCNNs through covalent functionalisation. Overall, we investigated the cytotoxicity of cisplatin in 2D monolayers of human glioma cell lines (U87) and 3D U87 tumour spheroid models.
Dissemination of the results:
Attendance at National and International conferences as planned has been limited due to the pandemic, but the Fellow has given 2 oral presentations at the 2022 UK and Ireland Controlled Release Society (UKICRS) Workshop & Symposium (in person) and 23rd symposium on "Signal Transduction at the Blood-Brain Barriers" (virtual) and a Poster Presentation and Small Talk at the 13th European and Global Summit for Nanomedicine", CLINAAM CLINAM 2022 Summit. The Fellow was also selected to give an oral presentation at the prestigious Controlled Release Society (CRS) 2022 Annual Meeting in Montreal, Canada but unfortunately due to pandemic her visa was not issued on time and no virtual meeting option was available at that conference.
As part of the MSCA project, the Fellow got involved in several public engagement activities like European Researchers’ Night (Month, 2020), Nottingham Soapbox Women in Science event (Month, 2022) about the importance of her MSCA project and spread her word to encourage women in STEM. In addition, the Fellow joined as a STEM ambassador West Midlands, UK, after starting my MSCA fellowship and took part in some of their public engagement activities. She also took part in some of the Patient and Public Involvement (PPI) meetings conducted by Trauma, Accident, Burns & Critical Care (TrABC) PPI Group meeting, and the PPI meeting organised by a BERTI, a brain tumour charity.
Preliminary result of the study has been published in Nano Energy (Arathyram Ramachandra Kurup Sasikala et al, Development of self-powered multifunctional piezomagnetic nanoparticles for non-invasive post-surgical osteosarcoma theranogeneration, Nano Energy, Volume 96, 1 June 2022, 107134) as open access publication in 2022. Moreover, further results from this project will be submitted as two manuscripts for peer review: one in Advanced Functional Materials and the other in Nano Energy as follows:
Arathyram Ramachandra Kurup Sasikala et al., BBB Penetrating Multifunctional Piezomagnetic Nanoneedles for Glioblastoma Treatment, [ to be submitted in Advanced Functional Materials by Mar 2023]
Arathyram Ramachandra Kurup Sasikala et al., Design and development of ultrasound responsive nanocarriers for cancer theranostics, [to be submitted in Nano Energy by Jan 2023]
The first research objective was to rationally design and characterise novel PMCNNs and this objective was met by the completion of WP1 by the preparation and characterisation of PMCNNs. For that first PiezoMagnetic nanoparticles (PMNPs) were fabricated and attached onto the f-CNTs through electrostatic and covalent functionalisation.
The second research objective was to evaluate the in vitro cytotoxicity of PMCNNs. This objective was met by the completion of WP2 by performing the toxicological evaluation of PMCNNs in different BBB cells Briefly the effect of concentration and incubation time of PMCNNs and their components were studied in vitro in different types of brain cells (human primary astrocytes and human brain microvascular endothelial cells etc.). Here we mainly used the modified lactate dehydrogenase (LDH) assay to circumvent the interference between carbon-based nanoparticles and fluorophores.
The third research objective was to investigate and elucidate the uptake mechanism(s) and fate of PMCNNs in BBB models. To achieve this objective, we successfully completed by assessing the translocation of PMCNNs in BBB models. Briefly a trans-well co-culture BBB model and multicellular spheroidal BBB model were developed to study the translocation of PMCNNs. The tightness of the BBB models was measured by imaging the expression of transmembrane proteins such as Claudin, Occludin and Zonula Occludens along with measuring the trans-endothelial electrical resistance (TEER) of the Transwell BBB models using an EVOM epithelial voltohmmeter. Cellular uptake of PMCNNs (25µg/ml) in BBB co-culture model were investigated in the presence and absence of ultrasound (US). The uptake of PMCNN was tracked through two photon microscopy imaging due to the intrinsic second harmonic generation imaging property of PMCNNs.
The fourth and final research objective was to evaluate the potential of PMCNNs as drug delivery system for the delivery of exemplar anticancer drugs in 3D Glioblastoma model. This objective was met by the completion of WP4 by evaluating the therapeutic potential of PMCNNs as delivery systems for anticancer therapeutics in 3D Glioblastoma Models. Briefly we assessed the therapeutic effects of PMCNNs as delivery systems for the anticancer drug cisplatin (CDDP) in 3D glioblastoma tumour spheroid models. Cisplatin (CDDP) was loaded onto PMCNNs through covalent functionalisation. Overall, we investigated the cytotoxicity of cisplatin in 2D monolayers of human glioma cell lines (U87) and 3D U87 tumour spheroid models.
Dissemination of the results:
Attendance at National and International conferences as planned has been limited due to the pandemic, but the Fellow has given 2 oral presentations at the 2022 UK and Ireland Controlled Release Society (UKICRS) Workshop & Symposium (in person) and 23rd symposium on "Signal Transduction at the Blood-Brain Barriers" (virtual) and a Poster Presentation and Small Talk at the 13th European and Global Summit for Nanomedicine", CLINAAM CLINAM 2022 Summit. The Fellow was also selected to give an oral presentation at the prestigious Controlled Release Society (CRS) 2022 Annual Meeting in Montreal, Canada but unfortunately due to pandemic her visa was not issued on time and no virtual meeting option was available at that conference.
As part of the MSCA project, the Fellow got involved in several public engagement activities like European Researchers’ Night (Month, 2020), Nottingham Soapbox Women in Science event (Month, 2022) about the importance of her MSCA project and spread her word to encourage women in STEM. In addition, the Fellow joined as a STEM ambassador West Midlands, UK, after starting my MSCA fellowship and took part in some of their public engagement activities. She also took part in some of the Patient and Public Involvement (PPI) meetings conducted by Trauma, Accident, Burns & Critical Care (TrABC) PPI Group meeting, and the PPI meeting organised by a BERTI, a brain tumour charity.
Preliminary result of the study has been published in Nano Energy (Arathyram Ramachandra Kurup Sasikala et al, Development of self-powered multifunctional piezomagnetic nanoparticles for non-invasive post-surgical osteosarcoma theranogeneration, Nano Energy, Volume 96, 1 June 2022, 107134) as open access publication in 2022. Moreover, further results from this project will be submitted as two manuscripts for peer review: one in Advanced Functional Materials and the other in Nano Energy as follows:
Arathyram Ramachandra Kurup Sasikala et al., BBB Penetrating Multifunctional Piezomagnetic Nanoneedles for Glioblastoma Treatment, [ to be submitted in Advanced Functional Materials by Mar 2023]
Arathyram Ramachandra Kurup Sasikala et al., Design and development of ultrasound responsive nanocarriers for cancer theranostics, [to be submitted in Nano Energy by Jan 2023]
The MSCA Fellowship has offered necessary support and funding for the development of the novel brain drug delivery system called PiezoMagnetic Carbon Nanoneedles and enabled the Fellow to take the next career step to independence by acquiring unique set of cutting-edge technological skills, knowledge, and transferable skills. Therefore, as part of the research we developed a smart and novel brain delivery system which has the ability to tackle neurological disorders non-invasively and ultimately extend and enhance the lives of patients affected by such conditions. The exciting result obtained from the project laid the foundations for future research and collaboration with the international scientific community, industry and all those interested in modulating the blood brain barrier, studying its biology and shuttling medicines to the brain.