Irisin is a myokine that stimulates browning of white adipocytes and enhances cortical bone mass during physical exercise. However, its short half-life limits its therapeutic potential, making sustained delivery systems desirable for bone tissue regeneration. In this study, we evaluated the effect of irisin-loaded porous silica nanoparticles (SiNPs) on caudal fin regeneration in a cholesterol-rich diet-induced obese medaka fish model, compared with irisin alone. Obese medaka were generated by feeding a 20% cholesterol diet for 2 months, followed by caudal fin amputation. Irisin formulations (free irisin or SiNP-loaded irisin at 350 ng and 700 ng) were administered via intraperitoneal injection. Cholesterol-fed medaka showed significant weight gain and fat accumulation over 30 days. Following fin amputation, bone regeneration was impaired in obese fish compared to normal controls. Notably, irisin-loaded SiNPs significantly enhanced fin regeneration, particularly at the higher dose (700 ng), compared to free irisin. These findings demonstrate that irisin delivered in porous silica nanoparticles improves bone regeneration in an obese medaka fish model more effectively than irisin alone.

Lipopolyplexes (LPPs), a hybrid nonviral vector that combines lipids and polymers with RNA or DNA, are emerging as a key platform for nucleic acid therapeutics such as mRNA vaccines, gene therapy, and CRISPR/Cas9. However, the influence of various lipid structures and their physicochemical characteristics on LPP formation is less well-understood. Here, we systematically evaluated how different lipid types affect cationic polyethylenimine (PEI)-based LPPs for delivering plasmid DNA (pDNA) encoding Cas9 endonuclease and single-guide RNA (sgRNA). LPPs formulated with four lipid types, cationic, ionizable, anionic, and neutral/fusogenic, and their combinations were evaluated for their influence on transfection efficiency and biocompatibility. Among the tested formulations, LPPs composed of anionic lipid Lecithin and fusogenic lipid DOPE exhibited excellent transfection efficiency and biocompatibility compared to those containing cationic lipid DOTAP and ionizable lipid DODMA. Further optimization with a small percentage of PEG lipid (DMG-PEG) resulted in a lead LPP formulation (Lecithin:Chol:DOPE:DMG-PEG) that achieved transfection efficiencies of 62.3%, 54.4%, and 52.6% in HEK293T, PANC1, and A549 cells, respectively, with significantly improved biocompatibility in vitro. The performance of this optimized LPP was significantly higher than that of the commercially available pDNA transfection reagent Lipofectamine 3000. By targeting the KRAS oncogene, the Lecithin:Chol:DOPE:DMG-PEG LPP system achieved gene-editing efficiencies of 18.3% and 15.6% in PANC1 and A549 cells, respectively, highlighting its therapeutic potential as a safe and effective nonviral delivery vehicle for CRISPR/Cas9-based gene editing and other nucleic acid delivery applications.

Immunocompromise is a hallmark of cancer, affecting both the peripheral immune system and local tumor microenvironment (TME). Current immunotherapies like checkpoint inhibitors, CAR-T cells, and neo-antigen vaccines show limited efficacy due to severe immunosuppression in most patients. Here, an immunologically engineered injectable immunehydrogel (iHG) is reported that can: i) recruit the desired set of immune cells away from the suppressed TME and peripheral organs, ii) activate them within a protective ambit of engineered immune-stimulatory hydrogel niche, and iii) release them to target cancer even in distant locations. Biodegradable and injectable iHG compositions are tested and optimized for their ability to attract and activate dendritic cells (DC), macrophages, monocytes, natural killer (NK) cells, B cells, and T cells via stimulator of interferon genes (STING), TLR, CD86, and Th1-polarized cytokine pathway without requiring exogenously introduced neo-antigens as vaccines. In a mouse melanoma model, optimized iHGs elicit a robust antitumor immune response through innate and adaptive arms. Most importantly, iHGs as a single agent immunomodulator exhibit better tumor control than when combined with anti-PD1 immune checkpoint antibody. These findings highlight the potential of engineering immunologically functional and injectable hydrogel niches as a new type of immunotherapeutics to reprogram immune cells to overcome both local and systemic immunosuppression and combat cancer effectively.

Simultaneous inhibition of multiple oncogenic signaling pathways is crucial for managing refractory cancers. This study introduces two unique core-shell nanoparticle (CS-NP) systems crafted from natural proteins that simultaneously target two crucial oncogenic pathways in refractory chronic myeloid leukemia (CML). Molecular analysis of approximately 14 refractory CML patients identified resistance to the standard treatment drug, imatinib, attributed to the overex-pression of the STAT5-transferrin pathway alongside the classic BCR-ABL fusion gene. To address this, we developed two dual-drug-loaded core-shell nanoparticles: (a) CS-NP1: Protamine sulfate nanocores carrying BCR-ABL siRNA and an albumin shell loaded with the STAT5 in-hibitor sorafenib, denoted as (PS-siRNA)-(Tf-Soraf) CS-NP; (b) CS-NP2: features a second-generation BCR-ABL inhibitor, dasatinib, in the albumin nanocore, and sorafenib in the transferrin nanoshell, labeled as (nAlb-Dasa)-(Tf-Soraf). We hypothesized that these dual-drug-loaded CS-NPs would effectively target both BCR-ABL and STAT5 pathways, with the transferrin nanoshell aiding in precise delivery to refractory CML cells overexpressing TfR1 due to STAT5 activity. Initial evaluations in drug resistant CML cell lines and patient-derived cells demonstrated significant cytotoxicity. Remarkably, even patients with BCR-ABL oncogene mutations displayed over 95% cytotoxicity with the CS-NPs. Furthermore, in vivo testing on a human xeno-graft model with a BCR-ABL+/+/STAT5+/+/TfR+/+ phenotype showcased a strong anti-tumor response. These results underscore the potential of a molecular-diagnosis-based rational design approach for protein-protein core-shell nanoparticles to simultaneously inhibit multiple oncogenic pathways, thereby overcoming resistance to targeted molecular therapies.

Abstract Successful cancer-immunotherapy need to address three critical factors: a) Relieving the immuno-suppressive tumor microenvironment (TME) b) expanding antigen specific, functionally avid effector T cells, c) triggering systemic immunity with a balanced innate-adaptive response for continued effector cell-feeder supply-chain and memory generation. Combinatorial engagement of such multi-elemental components demands smart, engineered immunotherapy systems compared to conventional vaccines. Here, we report an injectable Nano-Immuno-Gel (NIG) that can neutralize immunosuppressive components (e.g., myeloid derived suppressor cells (MDSCs), M2 macrophages (Mϕ) in TME, together with generating strong effector T cells by nano-vaccination, and sustained release of cytokines to enhance systemic immunity against tumor. In a proof-of-concept study, we show that sustained-release of myeloid-suppressive drug from NIG depot efficiently inhibited MDSCs and M2 Mϕ, not only in TME, but also in the circulation and in peripheral systems, thus creating a conducive, hot TME infiltrated with CD8 effector cells. Follow-up antigen vaccination with novel nano-adjuvant enhanced DC-T cell engagement and sustained IL15 release from NIG provided systemic NK/T cell activation, rendering significant control of the tumor growth. Our data demonstrate the potential of rationally engineered multi-functional nano-immuno-gels (NIG) to combine sequential neutralization of the suppressive TME, antigen vaccination, and concurrent systemic immune activation for highly effective combinatorial cancer immunotherapy. Citation Format: Anjali Kozhissery Sunilkumar, Margaux Salvi, Saira Nujoom Mohammed, Zahara Thaivalappil Zakkariya, Alejandra Gomez Cadena, Shalin Chappan, Sreedevi Panachithanathu Radhakrishnan, Maneesh Manohar, Pavithran Keechilat, Shantikumar Nair, Pedro Romero, Camilla Jandus, Manzoor Koyakutty. Combinatorial targeting of tumor-microenvironment together with antigen vaccination using innovative nano-immuno-gel for enhanced cancer-immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3735.

Tracking stem cell homing to the brain, their infiltration and brain tissue distribution, are critical for evaluating the efficacy of stem cell-based anti-glioma therapy. Here, we report the use of a biomineral magnetic nano-contrast agent, Fe-doped calcium phosphate nanoparticles (nCP:Fe), to label mesenchymal stem cells (MSCs) and track their migration towards brain tumor using magnetic resonance imaging (MRI). nCP:Fe labelled MSCs implanted intra-ventricularly in a rat C6-glioma model showed longitudinal migration towards tumor-region over 14 days, detected as hypointense signals in T2/T2* weighted-MR images, which was confirmed ex vivo using histological staining. Although, a small fraction of implanted MSCs migrated to the tumor-site, the study demonstrates that, MRI tracking of stem cells using nCP:Fe nano-contrast agent will facilitate better assessment of the efficacy of stem cell-guided anti-tumor therapy.

Remarkable progress has been made in the field of genome engineering after the discovery of CRISPR/Cas9 in 2012 by Jennifer Doudna and Emmanuelle Charpentier. Compared to any other gene-editing tools, CRISPR/Cas9 attracted the attention of the scientific community because of its simplicity, specificity, and multiplex editing possibilities for which the inventors were awarded the Nobel prize for chemistry in 2020. CRISPR/Cas9 allows targeted alteration of the genomic sequence, gene regulation, and epigenetic modifications using an RNA-guided site-specific endonuclease. Though the impact of CRISPR/Cas9 was undisputed, some of its limitations led to key modifications including the use of miniature-Cas proteins, Cas9 Retron precise Parallel Editing via homologY (CRISPEY), Cas-Clover, or development of alternative methods including retron-recombineering, Obligate Mobile Element Guided Activity(OMEGA), Fanzor, and Argonaute proteins. As cancer is caused by genetic and epigenetic alterations, gene-editing was found to be highly useful for knocking out oncogenes, editing mutations to regain the normal functioning of tumor suppressor genes, knock-out immune checkpoint blockade in CAR-T cells, producing ‘off-the-shelf’ CAR-T cells, identify novel tumorigenic genes and functional analysis of multiple pathways in cancer, etc. Advancements in nanoparticle-based delivery of guide-RNA and Cas9 complex to the human body further enhanced the potential of CRISPR/Cas9 for clinical translation. Several studies are reported for developing novel delivery methods to enhance the tumor-specific application of CRISPR/Cas9 for anticancer therapy. In this review, we discuss new developments in novel gene editing techniques and recent progress in nanoparticle-based CRISPR/Cas9 delivery specific to cancer applications.

Although tumor-antigen-based therapeutic cancer vaccines are a potential cancer immunotherapy strategy, recent clinical trials show low efficacy for multiple reasons. One method that has been recently investigated to improve the efficacy of therapeutic cancer vaccines is the development of implantable vaccines for sustained delivery of antigens and CD8 T cell activation. Here, we optimized the composition for an implantable vaccine scaffold composed of alginate, polyvinyl alcohol, and poly(methyl vinyl ether- alt-maleic anhydride) loaded with tumor antigens. Considering the adjuvant property of aluminum compounds, aluminum ion was used to crosslink alginate in the scaffold. The scaffold showed an effective antigen incorporation efficiency of 90.34 +/- 0.55% using ovalbumin as the model antigen and 89.67 +/- 2.8% using B16-F10 cell lysate. SEM analysis of the scaffold showed pore size ranging from 5 to 10 mu m. Cell viability analysis using mouse RAW 264.7 macrophages proved the cytocompatibility of the scaffold. In vitro antigen release studies using ovalbumin showed 8.42% release for a period of 14 days. In vivo antitumor analysis carried out in subcutaneous mouse B16-F10 melanoma model demonstrated that the scaffold vaccine reduced the rate of tumor growth and improved survival in tested animals. The median survival time increased from 29 days in untreated animals to 58 days in scaffold vaccine-implanted animals. Implantable scaffold vaccine against. Melanomaimage

Glioblastoma Multiforme (GBM) is one of the challenging tumors to treat as it recurs, almost 100%, even after surgery, radiation, and chemotherapy. In many cases, recurrence happens within 2-3cm depth of the resected tumor margin, indicating the inefficacy of current anti-glioma drugs to penetrate deep into the brain tissue. Here, we report an injectable nanoparticle-gel system, capable of providing deep brain penetration of drug up to 4 cm, releasing in a sustained manner up to >15 days. The system consists of ∼222 nm sized PLGA nanoparticles (NP-222) loaded with an anti-glioma drug, Carmustine (BCNU), and coated with a thick layer of polyethylene glycol (PEG). Upon release of the drug from PLGA core, it will interact with the outer PEG-layer leading to the formation of PEG-BCNU nanocomplexes of size ∼33 nm (BCNU-NC-33), which could penetrate >4 cm deep into the brain tissue compared to the free drug (< 5 mm). In vitro drug release showed sustained release of drug for 15 days by BCNU-NP gel, and enhanced cytotoxicity by BCNU-NC-33 drug-nanocomplexes in glioma cell lines. Ex vivo goat-brain phantom studies showed drug diffusion up to 4 cm in tissue and in vivo brain-diffusion studies showed almost complete coverage within the rat brain (∼1.2 cm), with ∼55% drug retained in the tissue by day-15, compared to only ∼5% for free BCNU. Rat orthotopic glioma studies showed excellent anti-tumor efficacy by BCNU-NP gel compared to free drug, indicating the potential of the gel-system for anti-glioma therapy. In effect, we demonstrate a unique method of sustained release of drug in the brain using larger PLGA nanoparticles acting as a reservoir while deep-penetration of the released drug was achieved by in situ formation of drug-nanocomplexes of size <50 nm which is less than the native pore size of brain tissue (> 100 nm). This method will have a major impact on a challenging field of brain drug delivery.

The success of a Raman spectroscopy device in cancer detection lies in its ability to acquire high‐quality Raman signals from samples and to employ efficient classification algorithms in analysing spectral data. Portable Raman systems enabled with artificial intelligence tools are well adaptable to clinical settings and for accuracy for community‐level rapid screening. Here, we developed a robotic Raman device with a high‐efficiency Raman probe, validating it against endometrial cancers detecting high‐grade, low‐grade cancers and normal classes. Algorithms like principal component analysis‐discriminant analysis, and support vector machine were compared against the deep learning methodology; convolutional neural network (CNN) with and without data augmentation. Eventually, the system could classify high‐grade, low‐grade and normal tissues with an F1‐score of 91%, 94% and 97%, respectively. CNN with data augmentation proved to be the most dependable classifier that works well even in the presence of high background noise. Thus, we demonstrate a unique portable Raman device with AI tools for high‐sensitivity Raman analysis of endometrial cancer.

Tumor microenvironment (TME) is a heterogeneous system consisting of both cellular and acellular components. The growth and progression of tumors rely greatly on the nature of TME, marking it as an important target in cancer immunotherapy. Lewis Lung Carcinoma (LLC) is an established murine lung cancer model representing immunologically 'cold' tumors characterized by very few infiltrated cytotoxic T-cells, high levels of Myeloid-Derived Suppressor Cells (MDSCs) and Tumor-Associated Macrophages (TAMs). Here, we report various strategies we applied to reverse the non-immunogenic character of this cold tumor by imparting: a) immunogenic cell death using Hypericin nanoparticle-based photodynamic therapy (PDT), b) repolarising TAM using a TLR7/8 agonist, resiquimod, c) immune checkpoint inhibition using anti-PD-L1 and d) depleting MDSCs using low-dose 5-fluorouracil (5-FU) chemotherapy. Interestingly, the nano-PDT, resiquimod or anti-PD-L1 treatment had no major impact on tumor growth, whereas low-dose 5-FU-mediated depletion of MDSCs showed significant anti-tumor effect, primarily caused by the increased infiltration of CD8+ cytotoxic T-cells (∼96%). Though we have tested combining PDT with resiquimod or 5-FU for any synergistic effect, low-dose 5-FU alone showed better response than combinations. In effect, we show that depletion of MDSCs using low-dose 5-FU was one of the best methods to augment infiltration of CD8+ cytotoxic T-cells into a cold tumor, which is resistant to conventional therapies including immune checkpoint inhibitors.

Immunotherapy is a promising approach in the management of human cancers and has been proven to provide a durable response in many cancers. It is helpful as an adjuvant therapy for cancers and at present is considered as a fourth pillar supporting surgery, chemotherapy and radiotherapy. In the treatment of oral cancer, immunotherapy is approved in late-stage diseases where surgical resection cannot be carried out or fails, leading to recurrences and metastasis. Evidences suggest that when given as a first-line treatment, it can elicit an immune response that shrinks tumours, which could provide long-term benefit for patients. But unlike the traditional approach which follows the uniform protocol for all oral cancer patients, effective immunotherapy requires a more site-specific personalized approach. The aim of this paper is to review the various immune evasive mechanisms adopted by tumour cells and their relevance as potential targets for immunotherapy in oral tongue squamous cell carcinoma.

The Central nervous system (CNS) control many critical functions such as intelligence, emotion, sense, memory, together with modulating many other functions of the entire human body. Diseases affecting CNS are challenging to treat because of the sensitive functional and structural features of CNS, and limited accessibility. Brain is secured from systemic circulation by a unique biological fence, the blood brain barrier (BBB), that limits molecular exchange between peripheral system and CNS. Most drugs used for treating CNS diseases fail to reach the brain in effective therapeutic concentrations due to the presence of BBB. During the last decade, specifically engineered drug delivery systems (DDS) have offered the opportunity to overcome systemic barriers in brain drug-delivery. However, even with the best engineered nano-carriers, delivery of sufficient dose of drug into the brain through systemic circulation remains a challenge. Here, we review various systems for ‘direct’ drug delivery to the brain using implantable or injectable intracranial nano-drug delivery systems (nDDS), which can ensure 100% release of drug to the brain, in a sustained or controlled fashion for prolonged periods, bypassing the BBB, thereby radically improving the therapeutic efficacy while minimizing systemic toxicity. Brain injectable nanogels or nanoparticle systems can be considered superior compared to other local drug-delivery systems because they possess better diffusivity of drugs. The nanogels can be injected through minimally invasive procedures, thereby precisely delivering drugs to specific target sites. This review covers the detailed discussion about locally injectable drug delivery systems and various nanogel based drug, oligonucleotide and theragnostic delivery systems used for treating brain malignancies, infections and neurodegenerative diseases.

The concept of hybrid drugs for targeting multiple aberrant pathways of cancer, by combining the key pharmacophores of clinically approved single-targeted drugs, has emerged as a promising approach for overcoming drug-resistance. Here, we report the design of unique hybrid molecules by combining the two pharmacophores of clinically approved BCR-ABL inhibitor (ponatinib) and HDAC inhibitor (vorinostat) and results of in vitro studies in drug-resistant CML cells. Robust 2D-QSAR and 3D-pharmacophore machine learning supervised models were developed for virtual screening of the hybrid molecules based on their predicted BCR-ABL and HDAC inhibitory activity. The developed 2D-QSAR model showed five information rich molecular descriptors while the 3D-pharmacophore model of BCR-ABL showed five different chemical features (hydrogen bond acceptor, donor, hydrophobic group, positive ion group, and aromatic rings) and the HDAC model showed four different chemical features (hydrogen bond acceptor, donor, positive ion group, and aromatic rings) for potent BCR-ABL and HDAC inhibition. Virtual screening of the 16 designed hybrid molecules identified FP7 and FP10 with better potential of inhibitory activity. FP7 was the most effective molecule with predicted IC50 using the BCR-ABL based 2D-QSAR model of 0.005 μM and that of the HDAC model of 0.153 μM, and that using the BCR-ABL based 3D-pharmacophore model was 0.02 μM and that with HDAC model was 0.014 μM. In vitro study (dose-response relationship) of FP7 in wild type and imatinib-resistant CML cell lines harboring Thr315Ile or Tyr253His mutations showed growth inhibitory IC50 values of 0.000 16, 0.0039, and 0.01 μM, respectively. This molecule also showed better biocompatibility when tested in whole blood and in PBMCs as compared to ponatinib or vorinostat.

Orally delivered molecularly targeted small-molecule drugs play a significant role in managing cancer as a chronic disease. However, due to the poor oral bioavailability of some of these molecules, high-dose administration is required leading to dose-limiting toxicity especially when delivered daily for a long duration. Here, we report an oral nanoformulation for small-molecule multi-kinase inhibitor, sorafenib tosylate, showing nearly two fold enhancement in the oral bioavailability and enhanced therapeutic efficacy with a better safety profile compared to the current clinical formulation. Using a scalable process involving high-pressure homogenization, sorafenib was loaded into an albumin nanocarrier at ~ 50 w/w%. Repeated preparation of gram-scale batches (n = 7) showed an average particle size of 180 ± 9 nm, encapsulation efficiency of 95 $$\pm$$ 2%, and drug-loading efficiency of 48 $$\pm$$ 0.7%. Further, surface engineering with a mucoadhesive layer on nanoparticles (referred to as ABSORF) resulted in the final size of 299 ± 38 nm and surface charge of −54 ± 8 mV. Single-dose and multidose pharmacokinetic studies showed two fold enhancement in the plasma concentration of sorafenib compared to current clinically used tablets. Antitumor efficacy studies in the orthotopic rat liver tumor model showed significant tumor regression (p value = 0.0037) even at half dose (eqv. to 200 mg of human equivalent dose) of ABSORF compared to clinical control (eqv. to 400 mg). The biodistribution of sorafenib from ABSORF was higher in the liver; however, liver and kidney function test parameters were comparable with that of the 2 × dose of clinical control. No abnormalities and signs of toxicity were seen in the histopathological analysis for ABSORF-treated animals. In summary, we demonstrate a scalable preparation of small-molecule drug-loaded nanoformulation with approximately two fold enhancement in oral bioavailability, improved antitumor efficacy, and acceptable toxicity profile.

Zoledronic acid (Zol) is a potent bisphosphonate drug used in cancer immunotherapy due to its ability to activate γδT cells and bring about depletion of tumor-associated macrophages. Zoledronic acid pulsed monocytes are utilized for the activation and proliferation of γδT cells. For adoptive γδT cell transfer, isolated peripheral blood mononuclear cells (PBMC) are treated with Zol under in vitro conditions for γδT cell expansion. Herein, we optimized, zoledronate conjugated calcium phosphate nanoparticles (Zol-nCP) to improve in vitro γδT cell proliferation. PBMC treated with Zol-nCP conjugates resulted in 28.7% enhancement in γδT cell proliferation in comparison to free Zol. In addition, the treatment of RAW 264.7 macrophages with Zol-nCP resulted in 25% enhanced Zol mediated toxicity compared to the bare drug. This enhancement in toxicity to macrophages can be utilized for better depletion of tumor-associated macrophages compared to free Zol. The development of this nanoconjugate may have great potential in enhancing the clinical efficacy of bisphosphonate-mediated cancer immunotherapy.

Tolerance induction is central to the suppression of autoimmunity. Here, we engineered the preferential uptake of nano-conjugated autoantigens by spleen-resident macrophages to re-introduce self-tolerance and suppress autoimmunity. The brain autoantigen, myelin oligodendrocyte glycoprotein (MOG), was conjugated to 200 or 500 nm silica nanoparticles (SNP) and delivered to the spleen and liver-resident macrophages of experimental autoimmune encephalomyelitis (EAE) mice, used as a model of multiple sclerosis. MOG-SNP conjugates significantly reduced signs of EAE at a very low dose (50 μg) compared to the higher dose (>800 μg) of free-MOG. This was associated with reduced proliferation of splenocytes and pro-inflammatory cytokines secretion, decreased spinal cord inflammation, demyelination and axonal damage. Notably, biodegradable porous SNP showed an enhanced disease suppression assisted by elevated levels of regulatory T cells and programmed-death ligands (PD-L1/2) in splenic and lymph node cells. Our results demonstrate that targeting nano-conjugated autoantigens to tissue-resident macrophages in lymphoid organs can effectively suppress autoimmunity.

Abstract Background Nanoparticle siRNA-conjugates are promising clinical therapeutics as indicated by recent US-FDA approval. In glioma stem cells (GSC), multiple stemness associated genes were found aberrant. We report intracranially injectable, multi-gene-targeted siRNA nanoparticle gel (NPG) for the combinatorial silencing of 3 aberrant genes, thus inhibiting the tumorogenic potential of GSCs. Methods NPG loaded with siRNAs targeted against FAK, NOTCH-1, and SOX-2 were prepared by the self-assembly of siRNAs with protamine–hyaluronic acid combination. Electron microscopy, DLS, and agarose gel electrophoresis were used for the physicochemical characterization. Cell transfection and gene-silencing efficiency were studied using human mesenchymal stem cells and rat C6 glioma-derived GSCs. Neurosphere inhibition was tested in vitro using GSCs derived from C6 cell line and glioma patient samples. Patient-derived xenograft model and orthotopic rat glioma model were used to test the effect of NPG on in vivo tumorigenicity. Results The siRNA nanoparticles with an average size ~ 250 nm and ~ 95% loading efficiency showed cellular uptake in ~95.5% GSCs. Simultaneous gene silencing of FAK, NOTCH-1, and SOX-2 led to the inhibition of neurosphere formation by GSCs, whereas normal stem cells remained unaffected and retained neuronal differentiation capability. GBM PDX models manifested significant impairment in the tumorigenic potential of NPG treated GSCs. Intracranial injection of NPG inhibited tumor growth in orthotopic rat brain tumor model. Conclusion Intracranially injectable n-siRNA NPG targeted to multiple stem-cell signaling impairs glioma initiation capabilities of GSCs and inhibited tumor growth in vivo.

This study investigated the adverse effects of 200 nm zinc oxide particles (nZnO) on sexual behavior and reproduction in Japanese medaka in comparison with ZnSO4 and correlated the consequences with the bioaccumulation pattern of the particles in associated organs. nZnO exposure impaired sexual and territorial behaviors and affected fertility by altering sperm viability and motility in males through reactive oxygen species (ROS) induction. Conversely, none of these effects other than behavior loss was seen in males exposed to ZnSO4. nZnO exposure to females induced ROS in ovaries, causing follicular growth arrest, atresia, and subfertility. Further, sex-steroid levels were altered by both nZnO and ZnSO4 in males and by nZnO but not ZnSO4 in females. Biodistribution studies revealed the deposition of nZnO as particulate matter in the brain, gills, gut, kidney, and ovary. Particle accumulation in the brain was sex specific, as the particles were found in the brain of males but not that of females. A similar trend was seen for zinc levels in males and females exposed to ZnSO4. Importantly, the female sex hormone, 17β-estradiol was found to prevent nZnO accumulation in the female brain, emphasizing the need for biodistribution profiling of nanoparticle-based drug delivery vehicles separately in males and females before they are commercialized. This study has demonstrated that the toxic effects of nZnO on the reproductive system were mainly caused by ROS induction, while zinc ions were predominantly responsible for the adverse impact of ZnSO4.

In oral squamous cell carcinoma (OSCC), expression of PDL1 is controversial with expressions showing a positive and negative correlation with survival in previous studies. Additionally, it is unclear whether expression on the tumour or tumour infiltrating lymphocytes (TIL) is a better predictor of survival. We performed this study on a cohort of Indian patients with OSCC to determine impact of PDL1 expression on survival. Retrospective analysis of 64 patients of OSCC treated with curative intent surgery with or without adjuvant therapy was performed. Stored tissue blocks were extracted and quantitative immunohistochemistry was performed for PDL1 expression separately on the tumour and the TIL using commercially available Dako kits. Correlation of clinical and pathological variables with PDL1 expression was performed using chi-square test. Survival analysis was performed using Kaplan-Meier method and Cox proportional hazards ratio. In our cohort, PDL1 expression was low, both in tumour (92% had <1% expression) and TIL (56% had <1% expression). Tumour low PDL1 expression (<1%) was associated with a higher risk of lymphovascular invasion (p = 0.044) and bone invasion (p = 0.01) but did not impact survival. Low TIL PDL1 expression (<1%) was more common in younger patients (<45 years) (p = 0.023) significantly predicting local recurrence (p = 0.02). PDL1 expression in OSCC was low. Low TIL PDL1 was common in younger patients and predicted local recurrence. Further study is required to better understand the relationship between age, tumour microenvironment and local recurrence.