Biology subjects

Shanker, A.

Publications and source records attributed to Shanker, A..

7 recordsLinked to original sources

bioRxiv2026

Engineering nanoparticle surface chemistry for antigen-presenting cell targeting improves specificity and safety of TLR3 agonist cancer immunotherapy

Innate immune agonists are promising therapeutic agents to induce immune responses against cancer. However, these agents have been limited by toxicity associated with systemic accumulation and activity in off-target cells. In this work, a targeted nanoparticle (NP) platform to encapsulate and protect the Toll-like receptor 3 (TLR3) agonist polyinosinic-polycytidylic acid (poly(I:C)) and promote its specific delivery to antigen presenting cells (APCs), macrophages and dendritic cells, for activation of this cell population was designed. To determine NP physiochemical properties that promote APC delivery, we developed a library of NP surface chemistries formed by electrostatic adsorption of polyanion coatings onto liposomes using layer-by-layer (LbL) assembly and screened the particles on APCs and off-target cells. Dextran sulfate was identified as a promising coating to enhance specific APC delivery. We applied these design parameters to develop a poly(I:C)-loaded NP for an APC-targeted immunotherapy. In a model of metastatic ovarian cancer, the LbL NP prolonged poly(I:C) retention in the peritoneal space--with 2-fold remaining 24-48hr after administration compared to free poly(I:C)--ultimately reducing systemic accumulation and associated toxicities. Compared to free drug, the NP reduced the increase in serum levels of TNF, IL-6, and CXCL10 by 9-, 4-, and 31-fold respectively. NP-treated mice experienced lower weight loss and recovered more quickly at a higher poly(I:C) dose, indicating a widening of the therapeutic window. The NP formulation enhanced accumulation of poly(I:C) in the tumor 2-fold and activation of the target APC population compared to free drug, and ultimately slowed tumor growth and extended survival in combination with doxorubicin chemotherapy. Overall, this work demonstrates a modular NP delivery strategy to improve the delivery, safety, and therapeutic window of a TLR3 agonist.

bioengineering↗

bioRxiv2025

Structure-Based Optimization of Pathogen Signal Sequences for Enhanced Antigen Expression in Humans for Vaccine Designs

Signal peptides (SPs) are short amino acid sequences found at the N-terminus of nascent polypeptides, which serve critical roles in trafficking, folding, and post-translational processing of mature proteins. Many sequence-based computational methods have been developed to predict and design optimal SPs for protein secretion in human cells. In this work, we introduce a structure-based workflow for identifying SPs and their regions, aiming to optimize SPs for protein secretion in human cells. Structural modeling of the SPs in complex with human signal recognition particle 54 kDa (SRP54), combined with hydrophobicity plots of the SPs and identification of the cleavage motif, can be used to detect SPs and SP regions. Afterwards, LigandMPNN is applied to redesign the SPs based on their structural complexes with SRP54. We employ our workflow to optimize the bacterial Yersinia pestis F1 SP, Lassa virus glycoprotein (GP) SP, and Venezuelan equine encephalitis virus (VEEV) E3 envelope protein for protein secretion in human cells to serve as vaccine candidates. By comparing the redesigned SPs with the original SPs, we propose that the binding affinity of SPs to SRP54 serve as the most important molecular determinant in the activity of the SPs. Notably, our experiments confirm that the structurally optimized SPs can express the Y. pestis F1 protein, Lassa GP, and VEEV GP in human cells. Overall, we demonstrate that structural modeling can serve as a valuable tool to predict the SPs and SP regions for vaccine antigens, predict whether the SPs can express mature proteins in humans, and optimize the SPs for the expression of mature proteins in humans.

biophysics↗

bioRxiv2025

SignalGen: A Protein Language Model Based AI Agent For Optimal Signal Peptide Prediction

Signal peptides are short amino acid sequences attached to the N-termini of mature proteins. They play determinant roles in protein expression as well as localization of mature proteins. While sequence-based machine learning (ML) models have been developed to identify the signal peptide sequences given the full or mature protein sequences, no model has been created to design optimal signal peptides with the localization of the mature proteins taken into account. Here, we develop a ML model that considers the mature protein sequence, organism, and localization as inputs, encodes and processes them through a Latent Residual Transformer (LRT), and outputs the optimal signal peptide sequences for enhanced expression of the mature proteins, regardless of whether the proteins are non-native to the organism or de novo. The model is trained using the latest data from the UniProt database up until July 2025. Benchmarking of our ML model shows good performance in predicting the signal peptides for both human and non-human proteins from the UniProt database. Furthermore, our ML model is implemented with an artificial intelligence (AI) agent to enhance accessibility for the general scientific community. Findings from this study provide a framework for predicting optimal signal peptides for non-native protein expression of viral and bacterial vaccine candidates in human cells and for enhanced expression of de novo proteins.

bioinformatics↗

bioRxiv2024

'Target-and-release' nanoparticles for effective immunotherapy of metastatic ovarian cancer

Immunotherapies such as checkpoint inhibitors (CPI) are effective in treating several advanced cancers, but these treatments have had limited success in metastatic ovarian cancer (OC). Here, we engineered liposomal nanoparticles (NPs) carrying a layer-by-layer (LbL) polymer coating that promotes their binding to the surface of OC cells. Covalent anchoring of the potent immunostimulatory cytokine interleukin-12 (IL-12) to phospholipid headgroups of the liposome core enabled the LbL particles to concentrate IL-12 in disseminated OC tumors following intraperitoneal administration. Shedding of the LbL coating and serum protein-mediated extraction of IL-12-conjugated lipids from the liposomal core over time enabled IL-12 to disseminate in the tumor bed following rapid NP localization in tumor nodules. Optimized IL-12 LbL-NPs promoted robust T cell accumulation in ascites and tumors in mouse models, extending survival compared to free IL-12 and remarkedly sensitizing tumors to CPI, leading to curative treatments and immune memory.

bioengineering↗

bioRxiv2024

Quantification of heterogeneity in human CD8+ T cell responses to vaccine antigens: an HLA-guided perspective

Vaccines have historically played a pivotal role in controlling epidemics. Effective vaccines for viruses causing significant human disease, e.g., Ebola, Lassa fever, or Crimean Congo hemorrhagic fever virus, would be invaluable to public health strategies and counter-measure development missions. Here, we propose coverage metrics to quantify vaccine-induced CD8+T cell-mediated immune protection, as well as metrics to characterize immuno-dominant epitopes, in light of human genetic heterogeneity and viral evolution. Proof-of-principle of our approach and methods will be demonstrated for Ebola virus, SARS-CoV-2, and Burkholderia pseudomallei (vaccine) proteins.

immunology↗

bioRxiv2020

Infra Red Thermography Reveals Transpirational Cooling in Pearl Millet (Pennisetum glaucum) Plants Under Heat Stress

An infra red thermographic analysis of well watered control and well watered heat stressed pearl millet (Pennisetum glaucum) was conducted at ICAR - Central Research Institute for Dryland Agriculture as a part of high resolution phenomics studies to identify the individual quantitative physiological parameters by plant phenotyping that form the basis for more complex abiotic stress tolerant traits. It was seen that the temperature gradient increased gradually from ground level to the top in the control non heat stressed plant. In contrast, it was seen that in the heat stressed plant the temperature increased up to the middle of the plant and then started to decrease at the top of the plant in comparison with the non heat stressed control plant. Our results indicate that the lowering of temperatures in the top of the heat stressed plant may be a mechanism by which the heat stressed plant acclimates to stress by regulating its transpiration thereby bringing in a cooling effect to counter stress.

plant biology↗

bioRxiv2019

Integrated high-resolution phenotyping, chlorophyll fluorescence induction kinetics and photosystem II dynamics under water stress and heat in Wheat (Triticum aestivum)

An experiment was conducted in controlled conditions in three varieties of wheat under water stress, heat and heat +water stress treatments with the objective of studying Chlorophyll a fluorescence, chlorophyll fluorescence induction kinetics and the function of Photosystem II by plant phenotyping as affected by stress. We hypothesised that during stress, specific adaptive strategies are employed by plants, such as structural and functional changes in PS II by which they acquire new homeostasis which may be protective adaptations. Water stress stress treatment was imposed on Water stress and Heat +Water stress treatments at 43 DAS. Heat treatment was imposed on 48 DAS. Maximum quantum yield of primary photochemistry was measured with PAM 2500 and OJIP was measured with FluorPen FP 100 after the onset of stress at four observation times on two days viz., pre-dawn and afternoon during stress. In addition continuous monitoring of photosynthetic efficiency was done with Monitoring PAM. Heat +Water stress stress was more detrimental as compared to Heat or Water stress alone in terms of maximum quantum yield of photochemistry. This could have been due to higher decrease in connectivity between PSII and its antennae resulting in lower photosynthetic efficiency resulting in the impairment and disruption of the electron transport. K step was observed in heat stress and heat +Water stress stress which may be because of damage to Oxygen Evolving Complex indicating that low thermostability of the complex. The stress treatments had a reduction in the plastoquinone pool size as indicated by the reduced area above the OJIP curve. Our study indicated that the instrument PAM 2500 sensed both stresses separately and combined earlier than the other instruments, so in terms of sensitivity PAM 2500 was more effective than FluorPen FP 100 and MultispeQ. Rapid screening of stress was more effectively with FluorPen FP 100 and MultispeQ than by PAM 2500.

plant biology↗