Qualification: 
Ph.D
anushaashokan@aims.amrita.edu

Dr. Anusha Ashokan earned her Bachelors (B. Tech) in Biotechnology and Biochemical Engineering from Mohandas College of engineering and Technology (Kerala University), her Masters (M. Tech) and PhD in Nanomedical Sciences from Amrita Centre for Nanosciences and Molecular Medicine (Amrita Vishwa Vidyapeetham). Her doctoral work focused on the development of multimodal contrast agents based on calcium phosphate nanoparticles and image guided therapeutics especially for MRI assisted radiofrequency ablation of liver tumor. After her PhD in 2014, she joined as a Research Scientist at Amrita Centre for Nanosciences and Molecular Medicine, where she focused on the translational challenges of calcium phosphate nanoparticle based therapeutics. In 2016, she received the DST Inspire Faculty Award and joined as DST Inspire Faculty at the Department of Biotechnology in Cochin University of Science and Technology. She is currently associated with ACNSMM as an Adjunct Faculty. Her current research areas include engineering of nanoparticles for cancer immunotherapy applications with specific focus on lymph node delivery and activation of a particular subset of T cells called γδT cells. Her achievements include SERB Early Career Research Award from Department of Science and Technology (2016), Young Scientist Travel Grant, CICS, Indian National Science Academy (2015), Senior Research Fellowship, Council of Scientific & Industrial Research (CSIR, 2012-2014), Travel Grant by Elsevier and A star Institute of Bioengineering and Nanotechnology (4th Nanotoday Conference, Dubai, 2015) and GATE Qualification (2007).

Research

Dr. Anusha’s lab focuses on the engineering of nanoparticles for different cancer immunotherapy applications. Lymph node targeted nanoparticles are being developed for effective activation of T cells against cancer by engineering its composition, size, charge and surface properties. In addition, we also focus on altering the biodistribution of immune modulating drugs for immunotherapy of cancer. Another major area of interest is the development of different strategies to activate a subset of T cells called γδ T cells against cancer. The lab is also involved in the translational work of calcium phosphate nanoparticle-based image guided therapeutics with a focus on MRI /CT assisted radiofrequency ablation of liver tumor.

Publications

Publication Type: Journal Article

Year of Publication Title

2019

Girish C. M., Nagareddy, R., Ramkumar, A., Krishnan, M., Harish, V., Anusha Ashokan, Shantikumar V. Nair, and Koyakutty, M., “Label-free Raman Signatures of Immune Cells: A New Tool for Artificial Intelligence in Immunotherapy”, Cancer Immunology Research, vol. 7, no. 2, p. B028, 2019.

2019

K. Meethaleve Sajesh, Anusha Ashokan, Gowd, G. Siddaraman, Dr. Manzoor K., Unni, A. K. K., and Shantikumar V Nair, “Magnetic 3D scaffold: A theranostic tool for tissue regeneration and non-invasive imaging in vivo.”, Nanomedicine: Nanotechnology, Biology, and Medicine, vol. 18, pp. 179-188, 2019.[Abstract]


We report an osteoconducting magnetic 3D scaffold using Fe doped nano-hydroxyapatite-Alginate-Gelatin (AGHFe1) for Magnetic Resonance Imaging based non-invasive monitoring of bone tissue regeneration. In rat cranial defect model, the scaffold facilitated non-invasive monitoring of cell migration, inflammatory response and matrix deposition by unique changes in transverse relaxation time (T2). Cell infiltration resulted in a considerable increase in T2 from ~37 to ~62 ms, which gradually returned to that of native bone (~23 ms) by 90 days. We used this method to compare in vivo performance of scaffold with bone-morphogenic protein-2 (AGHFe2) or faster degrading (AGHFe3). MRI and histological analysis over 90 days showed non-uniform bone formation in AGHFe1 with ∆T2 (T2 - T2 ) ~13 ms, whereas, AGHFe2 and AGHFe3 showed ∆T2 ~ 09 and 05 ms respectively, suggesting better bone formation in AGHFe3. Thus, we show that MR-contrast enabled scaffold can help better assessment of bone-regeneration non-invasively.

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2016

P. Sadanandan, Payne, N. L., Sun, G., Anusha Ashokan, Xavier, W., Gowd, S. G., Shantikumar V. Nair, Krishnakumar N. Menon, Bernard, C. C. A., and Koyakutty, M., “Nanoengineered Myelin Oligodendrocyte Glycoprotein Peptides Suppressed Experimental Autoimmune Encephalomyelitis: Implications In Multiple Sclerosis Therapy”, European Journal of Immunology, vol. 46 , 1 vol., pp. 316-317, 2016.

2016

V. H. Somasundaram, Pillai, R., Malarvizhi, G., Anusha Ashokan, Gowd, S., Peethambaran, R., Palaniswamy, S., Unni, A. K. K., Shantikumar V Nair, and Dr. Manzoor K., “Biodegradable Radiofrequency Responsive Nanoparticles for Augmented Thermal Ablation Combined with Triggered Drug Release in Liver Tumors”, ACS Biomaterials Science and Engineering, vol. 2, pp. 768-779, 2016.[Abstract]


Radiofrequency ablation (RFA) and doxorubicin (Dox) chemotherapy are separately approved for liver cancer therapy; however, both have limited success in the clinic due to suboptimal/nonuniform heating and systemic side effects, respectively. Here, we report a biodegradable nanoparticle (NP) system showing excellent RF hyperthermic response together with the ability to locally deliver Dox in the liver under RF trigger and control. The nanosystem was prepared by doping a clinically permissible dose (∼4.3 wt %, 0.03 ppm) of stannous ions in alginate nanoparticles (∼100 nm) coloaded with Dox at ∼13.4 wt % concentration and surface conjugated with galactose for targeting asialo-glycoprotein receptors in liver tumors. Targeted NP-uptake and increased cytotoxicity when combined with RF exposure was demonstrated in HEPG2 liver cancer cells. Following in vitro (chicken liver phantom) demonstration of locally augmented RF thermal response, in vivo scintigraphic imaging of 99Tc-labeled NPs was performed to optimize liver localization in Sprague-Dawley (SD) rats. RF ablation was performed in vivo using a cooled-tip probe, and uniformly enhanced (∼44%) thermal ablation was demonstrated with magnetic resonance imaging along with RF-controlled Dox release. In orthotopic rat liver tumor models, real-time infrared imaging revealed significantly higher (∼20 °C) RF thermal response at the tumor site, resulting in uniform augmented ablation (∼80%) even at a low RF power exposure of 15 W for just 1 min duration. Being a clinically acceptable, biodegradable material, alginate nanoparticles hold strong translational potential for augmented RF hyperthermia combined with triggered drug release.

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2015

N. Vadera, Anusha Ashokan, Genekehal S. Gowd, Sajesh, K. M., Chauhan, R. P., Dr. Jayakumar Rangasamy, Shantikumar V Nair, and Dr. Manzoor K., “Manganese doped Nano-bioactive Glass for Magnetic Resonance Imaging”, Materials Letters, vol. 160, pp. 335 - 338, 2015.[Abstract]


Magnetic resonance imaging (MRI) is an attractive method to image biomaterial implants owing to its high spatial resolution and absence of ionizing radiation. However, most of the biomaterials lack magnetic contrast sufficient enough to be imaged in MRI. Here, we report synthesis of manganese doped nano-bioactive glass (Mn-nBG) giving bright contrast for MRI. We have optimized a room temperature method of doping nBG for efficient T1 weighted magnetic contrast. In vitro study using primary mesenchymal stem cells showed no toxicity for Mn-nBG up to a tested concentration of 100µg/ml, suggesting potential applications in cell labeling and tissue engineering.

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2014

N. Ganesh, Anusha Ashokan, R kannan, R., Chennazhi, K., Shantikumar V Nair, and Dr. Manzoor K., “Magnetic Resonance Functional Nano-hydroxyapatite Incorporated Poly(caprolactone) Composite Scaffolds for in Situ Monitoring of Bone Tissue Regeneration by MRI”, Tissue Engineering - Part A, vol. 20, pp. 2783-2794, 2014.[Abstract]


In this study, we have reported the incorporation of a multi-modal contrast agent based on hydroxyapatite nanocrystals, within a poly(caprolactone)(PCL) nanofibrous scaffold by electrospinning. The multifunctional hydroxyapatite nanoparticles (MF-nHAp) showed simultaneous contrast enhancement for three major molecular imaging techniques. In this article, the magnetic resonance (MR) contrast enhancement ability of the MF-nHAp was exploited for the purpose of potentially monitoring as well as for influencing tissue regeneration. These MF-nHAp containing PCL scaffolds were engineered in order to enhance the osteogenic potential as well as its MR functionality for their application in bone tissue engineering. The nano-composite scaffolds along with pristine PCL were evaluated physico-chemically and biologically in vitro, in the presence of human mesenchymal stem cells (hMSCs). The incorporation of 30-40 nm sized MF-nHAp within the nanofibers showed a substantial increase in scaffold strength, protein adsorption, proliferation, and osteogenic differentiation of hMSCs along with enhanced MR functionality. This preliminary study was performed to eventually exploit the MR contrast imaging capability of MF-nHAp in nanofibrous scaffolds for real-time imaging of the changes in the tissue engineered construct. © Copyright 2014, Mary Ann Liebert, Inc.

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2013

Anusha Ashokan, Gowd, G. S., Somasundaram, V. H., Bhupathi, A., Peethambaran, R., Unni, A. K. K., ,, Shantikumar V Nair, and Dr. Manzoor K., “Multifunctional Calcium Phosphate Nano-contrast agent for Combined Nuclear, Magnetic and Near-Infrared Invivo Imaging”, Biomaterials, vol. 34, pp. 7143-7157, 2013.[Abstract]


Combination of three imaging techniques such as nuclear, magnetic and near-infrared fluorescence can aid in improved diagnosis of disease by synergizing specific advantages of each of these techniques such as deep tissue penetration of radiation signals, anatomical and functional details provided by magnetic contrast and better spatial resolution of optical signals. In the present work, we report the development of a multimodal contrast agent based on calcium phosphate nanoparticles (nCP), doped with both indocyanine green (ICG) and Gadolinium (Gd3+), and labeled with 99m-Technetium-methylene diphosphonate (99mTc-MDP) for combined optical, magnetic and nuclear imaging. In order to obtain the desired tri-modal contrast properties, the concentrations of ICG, Gd3+ and 99mTc were optimized at  0.15wt%, 3.38at% and  0.002ng/mg of nCP, respectively. The leaching-out of ICG was protected by an additional coating of polyethyleneimine (PEI). Toxicological evaluation of the final construct carried out on healthy human mononuclear cells, red-blood cells and platelets, showed excellent hemocompatibility. Invivo multimodal imaging using mice models revealed the ability to provide near-infrared, magnetic and nuclear contrast simultaneously. The nanoparticles also showed the potential for improved MR based angio-imaging of liver. Retention of intravenously administrated nanoparticles in the liver was reduced with PEGylation and the clearance was observed within 48h without causing any major histological changes in vital organs. Thus, we developed a non-toxic tri-modal nano-contrast agent using calcium phosphate nanoparticles and demonstrated its potential for combined nuclear, magnetic and near-infrared imaging invivo. © 2013 Elsevier Ltd.

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2012

Anusha Ashokan, Parwathy Chandran, Sadanandan, A. R., Chaitanya K. Koduri, Retnakumari, A. P., Menon, D., Shantikumar V Nair, and Dr. Manzoor K., “Development and Haematotoxicological Evaluation of Doped Hydroxyapatite based Multimodal Nanocontrast Agent for Near-infrared, Magnetic Resonance and X-ray Contrast Imaging”, Nanotoxicology, vol. 6, pp. 652-666, 2012.[Abstract]


Multimodal molecular imaging provides both anatomical and molecular information, aiding early stage detection and better treatment planning of diseased conditions. Here, we report development and nanotoxicity evaluation of a novel hydroxyapatite nanoparticle (nHAp) based multimodal contrast agent for combined near-infrared (NIR), MR and X-ray imaging. Under optimised wet-chemical conditions, we achieved simultaneous doping of nHAp (size ∼50 nm) with indocyanine green and Gd3+ contributing to NIR contrast (∼750–850 nm), paramagnetic behaviour and X-ray absorption suitable for NIR, MR and X-ray contrast imaging, respectively. Haematocompatibility studies using stem cell viability, haemolysis, platelet activation, platelet aggregation and coagulation time analysis indicated excellent compatibility of doped nHAp (D-nHAp). Further, the immunogenic function studies using human lymphocytes (in vitro) showed that D-nHAp caused no adverse effects. Collectively, our studies suggest that D-nHAp with excellent biocompatibility and multifunctional properties is a promising nanocontrast agent for combined NIR, MR and X-ray imaging applications

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2011

P. Chandran, Abhilash Sasidharan, Anusha Ashokan, Menon, D., Shantikumar V Nair, and Dr. Manzoor K., “Highly Biocompatible TiO2:Gd3+ Nano-contrast Agent with Enhanced Longitudinal Relaxivity for Targeted Cancer Imaging”, Nanoscale, vol. 3, pp. 4150-4161, 2011.[Abstract]


We report the development of a novel magnetic nano-contrast agent (nano-CA) based on Gd3+ doped amorphous TiO2 of size ∼25 nm, exhibiting enhanced longitudinal relaxivity (r1) and magnetic resonance (MR) contrasting together with excellent biocompatibility. Quantitative T1 mapping of phantom samples using a 1.5 T clinical MR imaging system revealed that the amorphous phase of doped titania has the highest r 1 relaxivity which is ∼2.5 fold higher than the commercially used CA Magnevist™. The crystalline (anatase) samples formed by air annealing at 250 °C and 500 °C showed significant reduction in r1 values and MR contrast, which is attributed to the loss of proton-exchange contribution from the adsorbed water and atomic re-arrangement of Gd 3+ ions in the crystalline host lattice. Nanotoxicity studies including cell viability, plasma membrane integrity, reactive oxygen stress and expression of pro-inflammatory cytokines, performed on human primary endothelial cells (HUVEC), human blood derived peripheral blood mononuclear cells (PBMC) and nasopharyngeal epidermoid carcinoma (KB) cell line showed excellent biocompatibility up to relatively higher doses of 200 μg ml-1. The potential of this nano-CA to cause hemolysis, platelet aggregation and plasma coagulation were studied using human peripheral blood samples and found no adverse effects, illustrating the possibility of the safe intravenous administration of these agents for human applications. Furthermore, the ability of these agents to specifically detect cancer cells by targeting molecular receptors on the cell membrane was demonstrated on folate receptor (FR) positive oral carcinoma (KB) cells, where the folic acid conjugated nano-CA showed receptor specific accumulation on cell membrane while leaving the normal fibroblast cells (L929) unstained. This study reveals that the Gd3+ doped amorphous TiO2 nanoparticles having enhanced magnetic resonance contrast and high biocompatibility is a promising candidate for molecular receptor targeted MR imaging. © 2011 The Royal Society of Chemistry.

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2010

Anusha Ashokan, Menon, D., Nair, S., and Dr. Manzoor K., “A Molecular Receptor Targeted, Hydroxyapatite Nanocrystal based Multi-modal Contrast Agent”, Biomaterials, vol. 31, pp. 2606-2616, 2010.[Abstract]


Multi-modal molecular imaging can significantly improve the potential of non-invasive medical diagnosis by combining basic anatomical descriptions with in-depth phenotypic characteristics of disease. Contrast agents with multifunctional properties that can sense and enhance the signature of specific molecular markers, together with high biocompatibility are essential for combinatorial molecular imaging approaches. Here, we report a multi-modal contrast agent based on hydroxyapatite nanocrystals (nHAp), which is engineered to show simultaneous contrast enhancement for three major molecular imaging techniques such as magnetic resonance imaging (MRI), X-ray imaging and near-infrared (NIR) fluorescence imaging. Monodispersed nHAp crystals of average size ∼30 nm and hexagonal crystal structure were in situ doped with multiple rare-earth impurities by a surfactant-free, aqueous wet-chemical method at 100 °C. Doping of nHAp with Eu3+ (3 at%) resulted bright near-infrared fluorescence (700 nm) due to efficient 5D0-7F4 electronic transition and co-doping with Gd3+ resulted enhanced paramagnetic longitudinal relaxivity (r1 ∼12 mM-1 s-1) suitable for T1 weighted MR imaging together with ∼80% X-ray attenuation suitable for X-ray contrast imaging. Capability of MF-nHAp to specifically target and enhance the signature of molecular receptors (folate) in cancer cells was realized by carbodiimide grafting of cell-membrane receptor ligand folic acid (FA) on MF-nHAp surface aminized with dendrigraft polymer, polyethyleneimine (PEI). The FA-PEI-MF-nHAp conjugates showed specific aggregation on FR+ve cells while leaving the negative control cells untouched. Nanotoxicity evaluation of this multifunctional nHAp carried out on primary human endothelial cells (HUVEC), normal mouse lung fibroblast cell line (L929), human nasopharyngeal carcinoma (KB) and human lung cancer cell line (A549) revealed no apparent toxicity even upto relatively higher doses of 500 μg/mL and 48 h of incubation. Flow-cytometry based reactive oxygen species (ROS) analysis also showed no significant levels of ROS generation in the nHAp treated cells. The tri-modal contrast imaging functionality together with molecular receptor targeting capability and biocompatibility makes MF-nHAp a promising biomineral contrast agent for combinatorial molecular imaging. © 2009 Elsevier Ltd. All rights reserved.

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Publication Type: Patent

Year of Publication Title

2017

Dr. Manzoor K., Anusha Ashokan, Harish, V., and Nair, S., “Radio-wave Responsive Doped Nanoparticles for Image Guided Therapeutics ”, U.S. Patent WO2017096342A1 (Filed)2017.

2017

Anusha Ashokan, Dr. Manzoor K., Shantikumar V Nair, M, S., and Harish, V., “MRI and CT Contrast-enabled Composite Implants for Image-guided Tissue Regeneration and Therapy (Filed)”, U.S. Patent PCT/US2017/0207932017.

2017

Anusha Ashokan, Dr. Manzoor K., Shantikumar V Nair, and Harish, V., “Non-Iodinated Radiolabeled Radiopaque Microbeads with MRI Contrast for Radioembolization (Filed)”, U.S. Patent PCT/US2017/0334862017.

2016

Dr. Manzoor K., Harish, V., Anusha Ashokan, and Nair, S., “Non-iodinated Radiopaque Microbeads, Capable of Binding to Radioisotopes for Trans-arterial Embolization and Trans-arterial Radioembolization”, U.S. Patent PCT No 201641017865 (Filed)2016.

2015

Dr. Manzoor K., Anusha Ashokan, Harish, V., and Shantikumar V Nair, “Radio-Wave Responsive Doped Nanoparticles for Image Guided Therapeutics (Filed)”, U.S. Patent 6495/CHE/20152015.

2012

Dr. Manzoor K., Anusha Ashokan, and Shantikumar V Nair, “THE ART, METHOD, MANNER, PROCESS AND SYSTEM OF NANO-BIOMINERAL FOR MULTIMODAL CONTRAST IMGING AND DRUG DELIVERY (Filed)”, U.S. Patent 25/CHE/20122012.

2009

Dr. Manzoor K., Anusha Ashokan, Menon, D., and Shantikumar V. Nair, “Multifunctional Nanobiomaterial for Molecular Imaging and Drug Delivery”, U.S. Patent # 2633/CHE/20092009.

2009

M. Koyakutty, Anusha Ashokan, Dr. Deepthy Menon, and Nair, S., “THE ART, METHOD, MANNER, PROCESS AND SYSTEM OF MULTIFUNCTIONAL NANOBIOMATERIAL FOR MOLECULAR IMAGING AND DRUG-DELIVERY (Filed)”, U.S. Patent 2633/CHE/20092009.