Therapeutic Effects of Ormeloxifene in Cervical Cancer Carcinogenesis

Cervical cancer (CxCa) remains the fourth leading cause of cancer related deaths among women worldwide. Cervical cancer is mainly (~ 99.7%) derived from high risk Human papillomavirus (HR HPV). HPV E6/E7 are the two main oncoproteins that interfere with p53 and pRb (retinoblastoma) cell cycle regulatory proteins and hinder their efficacy of controlling cell growth. Additionally, PI3K-Akt is a cell survival pathway that is aberrantly expressed in cervical cancer cells. This pathway has a profound role in inhibiting mitochondrial intrinsic apoptotic signaling pathway. Advanced stage cervical cancer is difficult to treat and patients diagnosed with metastatic disease have a poor survival rate. Therefore, there is an urgent need to develop newer treatment modalities. Ormeloxifene (ORM) is a non-hormonal, anti-estrogen, oral contraceptive for human use. Growing evidences also suggest that ormeloxifene has anti-cancerous properties in a variety of cancers. Developing nanoformulation of drugs has received much attention lately as nanoparticles have site specific targeted drug delivery. Nanoparticles have a specific size range that makes them capable of being entrapped and accumulated at the tumor site due to its leaky vasculature. As a result of it, drug is released from the particle core at a sustained rate; therefore, nanoparticulates offer enhanced bioavailability and better therapeutic efficacy. Considering these benefits, we engineered ormeloxifene loaded PLGA based novel nanoformulation (PLGA-ORM). In this work we validated anti-cancer properties of free ORM and its PLGA based nanoformulation. Our set of data showed that ormeloxifene significantly decreased the cellular proliferation and clonogenic potential of cervical cancer cells. Ormeloxifene also reduced the cellular motility and induced the apoptosis via targeting PI3K-Akt signaling in these cells. Furthermore, ormeloxifene modulated the HPV induced oncogenesis in Caski cells. Ormeloxifene also showed additive inhibitory effects on cellular proliferation and growth when used with radiation. Moreover, our novel PLGA-ORM had a particle size range of 100 – 280 nm and also exhibited excellent encapsulation of ormeloxifene in to PLGA core. PLGA-ORM was labeled with Coumarin 6 (green fluorescent) dye for its uptake studies, where PLGA-ORM internalized in cervical cancer cells in dose, time and energy dependent manner via endocytosis pathway. PLGA-ORM showed improved anti-proliferative/growth properties than free ormeloxifene in cervical cancer cells. When utilized in animals (an orthotopic mouse model) both ormeloxifene and PLGA-ORM showed great anti-tumorous properties, however PLGA-ORM had improved inhibitory effects on tumor growth than free ormeloxifene. To conclude, ormeloxifene and its nanoformulation have the potential to be a novel treatment modality for cervical cancer which can reduce the overall disease burden and improve patients’ life expectancy

miR-205: A Potential Biomedicine for Cancer Therapy

microRNAs (miRNAs) are a class of small non-coding RNAs that regulate the expression of their target mRNAs post transcriptionally. miRNAs are known to regulate not just a gene but the whole gene network (signaling pathways). Accumulating evidence(s) suggests that miRNAs can work either as oncogenes or tumor suppressors, but some miRNAs have a dual nature since they can act as both. miRNA 205 (miR-205) is one such highly conserved miRNA that can act as both, oncomiRNA and tumor suppressor. However, most reports confirm its emerging role as a tumor suppressor in many cancers. This review focuses on the downregulated expression of miR-205 and discusses its dysregulation in breast, prostate, skin, liver, gliomas, pancreatic, colorectal and renal cancers. This review also confers its role in tumor initiation, progression, cell proliferation, epithelial to mesenchymal transition, and tumor metastasis. Restoration of miR-205 makes cells more sensitive to drug treatments and mitigates drug resistance. Additionally, the importance of miR-205 in chemosensitization and its utilization as potential biomedicine and nanotherapy is described. Together, this review research article sheds a light on its application as a diagnostic and therapeutic marker, and as a biomedicine in cancer

A Study of Supervised Learning in Context with Decision and Regression Tree

One of the major objectives of machine learning is to instruct computers to use data or past experience to solve a specific task. Machine learning can be applied as association analysis through supervised learning, unsupervised learning but this paper will focus on strength of supervised learning classification algorithms. Section 1, describes the supervised learning with training sets. The function of Machine learning attempts is to tell system how to automatically find a good predictor based on past experiences. Section 2, deal with the supervised learning algorithm; the goal of supervised learning is to build a concise model of the distribution of class labels in terms of predictor features and analysis the decision tree and regression tree. Last section, describes the decision tree and regression tree and their results. The resulting classifier is then used to assign class labels to the testing instances where the values of the predictor features are known. The regression tree gives the value in term of continuous data

A Novel Exo-Glow Nano-system for Bioimaging

Background: Milk exosomes are widely used to improve the performance of various small macromolecules, oligonucleotides, and imaging agents for delivery and imaging applications. Indocyanine green (ICG) based Near-Infrared (NIR) fluorescent imaging is an attractive and safer technique used for number of clinical applications. However, ICG tend to have poor photostability, short half-life, non-specific proteins binding, and concentration-dependent aggregation. Therefore, there is an unmet clinical need to develop newer modalities to package and deliver ICG. Bovine milk exosomes are natural, biocompatible, safe, and feasible nanocarriers that facilitate the delivery of micro and macro molecules. Herein, we developed a novel exosomes based ICG nano imaging system that offers improved solubility and photostability of ICG. Methods: Following acetic acid based extracellular vesicles (EV) extraction method, we extracted the bovine milk exosomes from a variety of pasteurized fat-free milks. The EVs were screened for their physicochemical properties such as particle size and concentration, and zeta potential. Stability of these exosomes was also determined under different conditions including storage temperatures, pH, and salt concentrations. Next, ICG dye was loaded into these exosomes (Exo-Glow) via sonication method and further assessed for its fluorescence intensity and photostability using an IVIS imaging system. Results: Initial screening suggested that size of the selected bovine milk exosomes was from 100 - 135 nm with an average particle concentration of 5.8x102 particles/mL. Exo-Glow (ICG loaded exosomes) further showed higher fluorescence intensity of ~ 2x1010 MFI compared to free ICG (~ 8.1x109 MFI). Conclusions: These results showed that Exo-Glow has the potential to improve solubility, photostability, and biocompatibility of ICG and may serve as a safer NIR imaging tool for cells/tissues

Milk exosomes: Nature\u27s abundant nanoplatform for theranostic applications

Exosomes are a unique subpopulation of naturally occurring extracellular vesicles which are smaller intracellular membrane nanoparticle vesicles. Exosomes have proven to be excellent nanocarriers for carrying lipids, proteins, mRNAs, non-coding RNAs, and DNAs, and disseminating long-distance intercellular communications in various biological processes. Among various cell-line or biological fluid derived exosomes, milk exosomes are abundant in nature and exhibit many nanocarrier characteristics favorable for theranostic applications. To be an effective delivery carrier for their clinical translation, exosomes must inbuilt loading, release, targeting, and imaging/tracking characteristics. Considering the unmet gaps of milk exosomes in theranostic technology it is essential to focus the current review on drug delivery and imaging applications. This review delineates the efficiency of exosomes to load therapeutic or imaging agents and their successful delivery approaches. It is emphasized on possible modifications of exosomes towards increasing the stability and delivery of agents. This article also summarizes the specific applications and the process of developing milk exosomes as a future pharmaceutical drug delivery vehicle

Pluronic Polymer-Based Ormeloxifene Nanoformulations Induce Superior Anticancer Effects in Pancreatic Cancer Cells

Utilization of safe cytotoxic agents with precise anticancer activity is considered as the prime focus of cancer therapeutics research. A greater incentive for such agents arises from the molecules/drugs that are already being used for other indications. Ormeloxifene (ORM) is a nonsteroidal, nonhormonal selective estrogen receptor modulator (SERM), which has been in human use for contraception purposes. Although in the recent past, many reports have suggested its emerging role as an anticancer agent, no significant attention was paid toward generating simple and safe nanoformulation( s) for improved therapeutic activity and tumor cell-specific delivery. Our aim is to develop nanoformulation(s) of ormeloxifene to improve its targeted delivery in tumor cells. We developed ormeloxifene nanoformulation(s) by utilizing various biocompatible polymers. The optimized formulations with pluronic polymers F127 and F68 show improved nanoparticle characteristics. These formulations show enhanced cellular uptake that allows ormeloxifene’s intracellular availability. We further evaluated its improved anticancer activity by performing cell proliferation, flow cytometry, and immunoblotting assays. Overall, this study confirms possible novel nanoformulation(s) of ormeloxifene to be evolved as a new therapeutic modality for cancer treatment

Nebulization based Inhalation Nanomedicine for Lung Cancer Treatments

Background: Lung cancer is reported to have a high incidence rate and first leading cause of cancer-related morbidity and mortality across the world including in the United States. Noninvasive nebulized inhalation is a promising delivery strategy for lung, which can enhance the targeting efficiency and detention time interval of nanoparticles in the lung tissue, thus elevating the therapeutic index of therapeutic agent(s) at lower dosages. The aim of this study is to develop inhalable nanoparticles (INPs) for effective delivery of therapeutic agents in lung cancer cell lines and ex vivo models. Methods: The inhalation nanoparticles (INPs) were prepared by solvent evaporation and self-assembly approach. The INPs formulations were characterized by particle size, chemical composition, and drug loading efficiency using various analytical methods including FT-IR, DSC, SEM, and DSC/TGA. Cellular uptake of INPs was evaluated in 2D and 3D models of lung cancer cell lines (A549 and NCI-H1299) using fluorescence microscopy and flow cytometry analysis. Additionally, the therapeutic evaluation of gambogic acid and gemcitabine encapsulated INPs was performed by basic in vitro biological assays using proliferation (CCK-8), mucoadhesion Boyden chamber, and apoptosis assays using lung cancer (A549 and NCI-H1299) monolayers, spheroids, and xenograft tumors. Results: The developed INPs exhibited an average size of ~110 nm in dynamic light scattering measurements. INPs formulation showed a remarkable mucoadhesion and mucopenetration potential in-vitro model(s). Cellular uptake studies demonstrated that INPs formulation facilitates an effective endosomal release into the cytosol. The in vitro study confirms that INPs release the drugs in a sustained manner. Additionally, the INPs formulation showed superior in vitro anti-cancer activity in lung cancer cell lines, spheroids and xenograft tumor. Conclusions: Altogether this study confirms that INPs formulation demonstrates an improved therapeutic benefit over free drug against lung cancer cell lines, spheroids and xenograft tumor. This study could lead as an innovative therapeutic modality for the treatment of lung cancer

A Novel Exo-Glow Nano-system for Cellular Imaging

Background: Indocyanine green (ICG) based Near-Infrared (NIR) fluorescent imaging is an attractive and safer technique used for number of clinical applications. However, ICG tend to have poor photostability, short half-life, non-specific proteins binding, and concentration-dependent aggregation. Therefore, there is an unmet clinical need to develop newer modalities to package and deliver ICG. Bovine milk exosomes are natural, biocompatible, safe, and feasible nanocarriers that facilitate the delivery of micro and macro molecules. Herein, we developed a novel exosomes based ICG nano imaging system that offers improved solubility and photostability of ICG. Methods: Following acetic acid based extracellular vesicles (EV) extraction method, we extracted the bovine milk exosomes from a variety of pasteurized fat-free milks. The EVs were screened for their physicochemical properties such as particle size and concentration, and zeta potential. Stability of these exosomes was also determined under different conditions including storage temperatures, pH, and salt concentrations. Next, ICG dye was loaded into these exosomes (Exo-Glow) via sonication method and further assessed for its fluorescence intensity and photostability using an IVIS imaging system. Results: Initial screening suggested that size of the selected bovine milk exosomes was from 100 - 135 nm with an average particle concentration of 5.8x102 particles/mL. Exo-Glow (ICG loaded exosomes) further showed higher fluorescence intensity of ~ 2x1010 MFI compared to free ICG (~ 8.1x109 MFI). Conclusions: These results showed that Exo-Glow has the potential to improve solubility, photostability, and biocompatibility of ICG and may serve as a safer NIR imaging tool for cells/tissues