Single-cell transcriptional signature-based drug repurposing and in vitro evaluation in colorectal cancer.

BACKGROUND: The need for intelligent and effective treatment of diseases and the increase in drug design costs have raised drug repurposing as one of the effective strategies in biomedicine. There are various computational methods for drug repurposing, one of which is using transcription signatures, especially single-cell RNA sequencing (scRNA-seq) data, which show us a clear and comprehensive view of the inside of the cell to compare the state of disease and health. METHODS: In this study, we used 91,103 scRNA-seq samples from 29 patients with colorectal cancer (GSE144735 and GSE132465). First, differential gene expression (DGE) analysis was done using the ASAP website. Then we reached a list of drugs that can reverse the gene signature pattern from cancer to normal using the iLINCS website. Further, by searching various databases and articles, we found 12 drugs that have FDA approval, and so far, no one has reported them as a drug in the treatment of any cancer. Then, to evaluate the cytotoxicity and performance of these drugs, the MTT assay and real-time PCR were performed on two colorectal cancer cell lines (HT29 and HCT116). RESULTS: According to our approach, 12 drugs were suggested for the treatment of colorectal cancer. Four drugs were selected for biological evaluation. The results of the cytotoxicity analysis of these drugs are as follows: tezacaftor (IC10 = 19 µM for HCT-116 and IC10 = 2 µM for HT-29), fenticonazole (IC10 = 17 µM for HCT-116 and IC10 = 7 µM for HT-29), bempedoic acid (IC10 = 78 µM for HCT-116 and IC10 = 65 µM for HT-29), and famciclovir (IC10 = 422 µM for HCT-116 and IC10 = 959 µM for HT-29). CONCLUSIONS: Cost, time, and effectiveness are the main challenges in finding new drugs for diseases. Computational approaches such as transcriptional signature-based drug repurposing methods open new horizons to solve these challenges. In this study, tezacaftor, fenticonazole, and bempedoic acid can be introduced as promising drug candidates for the treatment of colorectal cancer. These drugs were evaluated in silico and in vitro, but it is necessary to evaluate them in vivo.

Core decompression combined with local DFO administration loaded on polylactic glycolic acid scaffolds for the treatment of osteonecrosis of the femoral head: a pilot study.

BACKGROUND: Deferoxamine (DFO) angiogenesis induction potential has been demonstrated in earlier studies, but not in the osteonecrosis of the femoral head (ONFH). In this study, we evaluated the outcome of ONFH treated with combined core decompression and local DFO administration loaded on Polylactic Glycolic Acid (PLGA). PATIENTS AND METHODS: In a pilot experimental study, six patients (10 hips) with early-stage non-traumatic ONFH were treated by core decompression, and concurrent injection of local DFO loaded on PLGA scaffold into the subchondral femoral head. Outcome measures were evaluated before the surgery and 12 and 24 months after the surgery and included visual analog scale (VAS) for pain, modified Merle d'Aubigné-Postel (MAP) score for hip function by MRI, and rate of osteonecrosis assessed by the modified. RESULTS: The mean MPA score was 14.7 ± 1.16 before the surgery and 16.7 ± 1.41 one year after the surgery (P = 0.004). The mean VAS for pain was 4.7 ± 1.25 before the surgery and 1.8 ± 1.03 one year after the surgery (P = 0.005). The mean Kerboul angle was 219 ± 58.64 before the operation and 164.6 ± 41.82 one year after the operation (P < 0.001). Osteonecrosis progression or collapse was not seen in any of the patients at the final follow-up. No postoperative side effect attributed to the DFO was noticed, as well. CONCLUSION: In short-term follow-up, combined core decompression and local DFO administration not only prevent the progression of ONFH but also reduces the rate of osteonecrosis significantly. However, future controlled studies are required to confirm the present results. TRIAL REGISTRATION: IRCT20161121031003N3, 16/04/2019.

Engineered mesoporous silica nanoparticles, new insight nanoplatforms into effective cancer gene therapy.

The use of nucleic acid to control the expression of genes relevant to tumor progression is a key therapeutic approach in cancer research. Therapeutics based on nucleic acid provide novel concepts for untreatable targets. Nucleic acids as molecular medications must enter the target cell to be effective and obstacles in the systemic delivery of DNA or RNA limit their use in a clinical setting. The creation of nucleic acid delivery systems based on nanoparticles in order to circumvent biological constraints is advancing quickly. The ease of synthesis and surface modification, biocompatibility, biodegradability, cost-effectiveness and high loading capability of nucleic acids have prompted the use of mesoporous silica nanoparticles (MSNs) in gene therapy. The unique surface features of MSNs facilitate their design and decoration for high loading of nucleic acids, immune system evasion, cancer cell targeting, controlled cargo release, and endosomal escape. Reports have demonstrated successful therapeutic outcomes with the administration of a variety of engineered MSNs capable of delivering genes to tumor sites in laboratory animals. This comprehensive review of studies about siRNA, miRNA, shRNA, lncRNA and CRISPR/Cas9 delivery by MSNs reveals engineered MSNs as a safe and efficient system for gene transfer to cancer cells and cancer mouse models.

Bicondylar Hoffa fracture with concurrent medial and lateral collateral ligament avulsion: A case report.

Hoffa fractures are rare fractures of the femoral condyle that occur in the coronal plane of the bone. In most cases, high-energy trauma leads to isolated coronal fractures of one of the femoral condyles, medial or lateral. Even with a typical unicondylar Hoffa fracture, our patient sustained a bicondylar Hoffa fracture in his right knee after falling from high and suffering direct trauma as well. The fracture was approached from both the medial and lateral sides of the distal femur. Three-month follow-up showed excellent functional scores, no laxity, and no pain.

Co-Delivery of Silymarin and Metformin Dual-Loaded in Mesoporous Silica Nanoparticles Synergistically Sensitizes Breast Cancer Cell Line to Mitoxantrone Chemotherapy.

The development nano-carriers based therapeutic methods is a potent strategy for enhancing cellular delivery of drugs and therapeutic efficiency in cancer chemotherapy. In the study, silymarin(SLM) and metformin (Met) were co-loaded into mesoporous silica nanoparticles (MSNs) and evaluated the synergistic inhibitory effect of these natural herbal compound in improving chemotherapeutic efficiency against MCF7MX and MCF7 human breast cancer cells. Nanoparticles have been synthesized and characterized by FTIR, BET, TEM, SEM, and X-ray diffraction. Drug loading capacity and release determined. The both single and combined form of SLM and Met (free and loaded MSN) were used for MTT assay, colony formation and real time-PCR in cellular study. The synthesis MSN were uniformity in size and shape with particle size of approximately 100 nm and pore size of approximately 2 nm. The Met-MSNs IC30, SLM -MSNs IC50 and dual-drug loaded MSNs IC50 were much lower than of free-Met IC30, free-SLM IC50 and free Met-SLM IC50 MCF7MX and MCF7cells. The co-loaded MSNs treated cells were increased sensitivity to mitoxantrone with the inhibition of BCRP mRNA expressions and could induce apoptosis in MCF7MX and MCF7 cells in comparison with other groups. Colony numbers were significantly reduced in comparison to with other groups in the co-loaded MSNs -treated cells ( ). Our results indicate that Nano-SLM enhances the anti-cancer effects of SLM against human breast cancer cells. The findings of the present study suggest that the anti-cancer effects of both metformin and silymarin enhances against breast cancer cells when MSNs are used as a drug delivery system.

Mechanism and function of miR-140 in human cancers: A review and in silico study.

MicroRNA-140 (miR-140) acts as a tumor suppressor and plays a vital role in cell biological functions such as cell proliferation, apoptosis, and DNA repair. The expression of this miRNA has been shown to be considerably decreased in cancer tissues and cell lines compared with normal adjacent tissues. Consequently, aberrant expression of some miR-140 target genes can lead to the initiation and progression of various human cancers, such as breast cancer, gastrointestinal cancers, lung cancer, and prostate cancer. The dysregulation of the miR-140 network also affects cell proliferation, invasion, metastasis, and apoptosis of cancer cells by affecting various signaling pathways. Besides, up-regulation of miR-140 could enhance the efficacy of chemotherapeutic agents in different cancer. We aimed to cover most aspects of miR-140 function in cancer development and address its importance in different stages of cancer progression.

Association of CT Scan Parameters with the Risk of Renal Angiomyolipoma Rupture; a Brief Report.

INTRODUCTION: Rupture of renal angiomyolipoma (AML) is an emergency and life-threatening complication. This study aimed to evaluate the association of computed tomography (CT) scan parameters with the risk of rupture in renal AMLs. METHODS: In this retrospective cross-sectional study, patients who were referred to a referral university hospital with diagnosis of AML, between 2007 and 2019, were included. Patients were divided into ruptured and non-ruptured cases based on surgery and CT scan findings and the baseline characteristics as well as CT scan parameters were compared between the two groups. RESULTS: 20 AML patients with the mean age of 39.6 ± 12.5 years were included (75% female). The lesion was ruptured in 8 (40%) patients. The mean size of the lesion was 97.0 ± 15.9 mm in the ruptured and 72.0 ± 29.4 in the non - ruptured AML ( p = 0.045). The mean fat density based on non-contrast enhanced CT (NCCT) scan (- 56.1 ± 16.3 vs - 74.9±24.1; p = 0.018) and contrast enhanced CT (CECT) scan (- 20.8 ± 16.9 vs - 50.5 ± 31.7; p = 0. 016) was significantly higher in the ruptured cases. Total tumor density based on NCCT scan was significantly greater in the ruptured AMLs ( 19.6 ± 25.9 vs -22.7±41.6, p=0.033). CONCLUSION: It seems that some CT scan parameters such as mean fat density and total tumor density could be used for differentiation between ruptured and non-ruptured AMLs.

siRNA delivery using intelligent chitosan-capped mesoporous silica nanoparticles for overcoming multidrug resistance in malignant carcinoma cells.

Although siRNA is a promising technology for cancer gene therapy, effective cytoplasmic delivery has remained a significant challenge. In this paper, a potent siRNA transfer system with active targeting moieties toward cancer cells and a high loading capacity is introduced to inhibit drug resistance. Mesoporous silica nanoparticles are of great potential for developing targeted gene delivery. Amino-modified MSNs (NH2-MSNs) were synthesized using a modified sol-gel method and characterized by FTIR, BET, TEM, SEM, X-ray diffraction, DLS, and 1H-NMR. MDR1-siRNA was loaded within NH2-MSNs, and the resulting negative surface was capped by functionalized chitosan as a protective layer. Targeting moieties such as TAT and folate were anchored to chitosan via PEG-spacers. The loading capacity of siRNA and the protective effect of chitosan for siRNA were determined by gel retardation assay. MTT assay, flow cytometry, real-time PCR, and western blot were performed to study the cytotoxicity, cellular uptake assay, targeting evaluation, and MDR1 knockdown efficiency. The synthesized NH2-MSNs had a particle size of ≈ 100 nm and pore size of ≈ 5 nm. siRNA was loaded into NH2-MSNs with a high loading capacity of 20% w/w. Chitosan coating on the surface of siRNA-NH2-MSNs significantly improved the siRNA protection against enzyme activity compared to naked siRNA-NH2-MSNs. MSNs and modified MSNs did not exhibit significant cytotoxicity at therapeutic concentrations in the EPG85.257-RDB and HeLa-RDB lines. The folate-conjugated nanoparticles showed a cellular uptake of around two times higher in folate receptor-rich HeLa-RDB than EPG85.257-RDB cells. The chitosan-coated siRNA-NH2-MSNs produced decreased MDR1 transcript and protein levels in HeLa-RDB by 0.20 and 0.48-fold, respectively. The results demonstrated that functionalized chitosan-coated siRNA-MSNs could be a promising carrier for targeted cancer therapy. Folate-targeted nanoparticles were specifically harvested by folate receptor-rich HeLa-RDB and produced a chemosensitized phenotype of the multidrug-resistant cancer cells.

The effect of oleuropein on apoptotic pathway regulators in breast cancer cells.

In spite of advancements in breast cancer therapy, this disease is still one of the significant causes of women fatalities globally. Dysregulation of miRNA plays a pivotal role in the initiation and progression of cancer. Therefore, the administration of herbal compounds with anticancer effects through controlling microRNA expression can be considered as a promising therapy for cancer. Oleuropein is the most prevalent phenolic compound in olive. Given its domestic consumption, low cost, and nontoxicity for human beings, oleuropein can be used in combination with the standard chemotherapy drugs. To this end, we examined the effect of oleuropein on two breast cancer cell lines (MCF7 and MDA-MB-231). Our findings revealed that oleuropein significantly decreased cell viability in a dose- and time-dependent manner, while it increased the apoptosis in MCF7 and MDA-MB-231 cells. In the presence of oleuropein, the expression levels of miR-125b, miR-16, miR-34a, p53, p21, and TNFRS10B increased, while that of bcl-2, mcl1, miR-221, miR-29a and miR-21 decreased. The findings pointed out that oeluropein may induce apoptosis via not only increasing the expression of pro-apoptotic genes and tumor suppressor miRNAs, but also decreasing the expression of anti-apoptotic genes and oncomiR. Consequently, oleuropein can be regarded as a suitable herbal medication for cancer therapy.

Engineering mesenchymal stem cells: a novel therapeutic approach in breast cancer.

Breast cancer is one of the most prevalent and deadliest cancers among women in the world because of its aggressive behaviour and inadequate response to conventional therapies. Cellular and gene therapies based on mesenchymal stem cells (MSCs) represent promising treatment strategies for multiple diseases, such as cancers. MSCs are multipotent adult stem cells with important features for cell therapy, such as tissue homing to injured sites, their differentiation potential, their capacity of secreting plenty of trophic factors, and low immunogenicity. The quite easy isolation of these cells from various types of tissues are associated with no ethical concern when dealing with foetal or embryonic stem cells. The MSCs exhibit both pro and anti-oncogenic properties. However, genetic engineering of MSCs and nanoparticles is being employed as a means to solve some of these problems and improve the antitumor properties of these cells. The tumour-homing ability of MSCs and their exosomes to tumour niches have made them as a promising vector for targeted delivery of therapeutic agents to tumours site. The present study investigated MSCs specifications, pro- and anti-oncogenic properties of MSCs in breast cancer, and reviewed targeted breast cancer therapy via engineered MSCs, likely as potent cellular vehicles.

Design and evaluation of biological gate circuits and their therapeutic applications in a model of multidrug resistant cancers.

OBJECTIVES: Synthetic biology is primarily an emerging research field that consists of designing new synthetic gene circuits dedicated to targeted functions and therapies such as cancer therapy. In this study, a genetic logic NOT-IF gate is used to reduce the multidrug resistance and facilitate the malignant cancer therapy. MCF7 cancer cells were cultured in RPMI-1640 medium and transfected with lentiviral vectors including MDR1 gene and the corresponding shRNA against MDR1 with controllable promoters. Transcript levels and protein levels of MDR1 gene were quantified. RESULTS: Our results showed that when doxycycline (DOX) and sodium butyrate were present and IPTG was absent, these led to a 74,354-fold increase in MDR1 gene expression. Upon IPTG treatment, the MDR1 gene expression was not detected due to the lack of the inducer. In addition, following IPTG induction in the presence of DOX and sodium butyrate and expressing shRNA, there was a 75% reduction in MDR1 gene expression compared to those cells treated only with sodium butyrate and DOX. CONCLUSIONS: We successfully designed and implemented the genetic logic NOT-IF gate at the transcriptional level using the inducible expression of both MDR1 drug resistance pump and its specific shRNA in MCF7 cancer cells, using the third generation lentiviral vectors.

Genetically modified Pichia pastoris, a powerful resistant factory for gold and palladium bioleaching and nanostructure heavy metal biosynthesis.

A metal-resistant engineered Pichia pastoris was developed here to fulfil the metal bioleaching in aqueous conditions. Parent and recombinant yeasts were grown in YPD medium containing different concentrations of ion metals. XRD, electron microscopy and particle size analyser were used for the characterisation and the nanoparticle analyses. The nanoparticle production kinetics were studied by ICP-OES. The cytotoxicity of nanoparticles was assayed against human cell lines. Media colours changed to a range from purplish-brown to grey during early fermentation stages. The maximum biosorption capacities were recorded 81.23 and 493.35 mg/g for gold and palladium in batch conditions, respectively. Various physical investigations proved monodispersed spherical nanoparticles around 100 nm in size. Pure palladium nanoparticles and PdCl2 represented the least cytotoxic potency towards T47D and EPG85.257 cells. The results demonstrated that the genetically modified yeast is a cost-effective, high-throughput, robust, and facile system for metal biosorption.

Synthetic Biology Based on Genetic Logic Circuit, Using the Expression of Drug Resistance, BCRP Pump in MCF-7 Cancer Cell Line.

Biological circuits are developed as biological parts within a cell to carry out logical functions resembling those studied in electronics circuits. These circuits can be performed as a method to vary cellular functions, to develop cellular responses to environmental conditions, or to regulate cellular developments. This research explored the possibility of synthetic biology based on the genetic logic circuit A and (not B) using the inducible expression of the both BCRP drug resistance pump and its specific shRNA in MCF-7 cancer cell line utilizing the third generation of lentiviral vectors. The accuracy of the output of the proposed circuit for living cells, was confirmed by the results of the Real-Time PCR and flow cytometry at the RNA and protein levels. At the RNA level, the effect of the inducers on the BCRP gene expression and silencing were investigated by real-time PCR. Furthermore, at the protein level, induction of the expression of the BCRP pump resulted in driving out of the substrate from inside the cells leading to the decrease of the fluorescent emission from the transfected cells. We successfully designed and implemented the genetic logic circuit A and (not B) using the inducible expression of the both BCRP drug resistance pump and its specific shRNA in MCF-7 cancer cells.

Antispasmodic Effects of Aqueous and Hydroalcoholic Punica granatum Flower Extracts on the Uterus of Non-pregnant Rats.

BACKGROUND: Punica granatum Linn. (PG) is native to the Mediterranean region. Its flower exhibited antioxidant activity. The present study attempt to investigate the effect of these extract on uterine contraction and its possible mechanism(s). METHODS: Thirty five female Wistar rats (200-300 g) at estrous phases of cycle was examined in this study; pieces of virgin adult rat uterus (1.5 cm) were suspended in an organ bath containing 10 ml of De Jalon solution at 29 °C. Tissue contractility was isometrically recorded. KCl (60 mM), BaCl2 (4 mM) and oxytocin (10 mU/ml) were applied to the tissue in the presence and absence of aqueous and hydroalcoholic extracts of the plant (0.05, 0.1, 0.2, 0.4 and 0.8 mg/ml). Propranolol (1 µM) and naloxane (1 µM) were added in KCl induced contractions. The results were analyzed by ANOVA and p < 0.05 were considered as significant. RESULTS: Cumulative concentration of extracts reduced uterine contractions induced by KCl dose-dependently (p < 0.01). Extracts in a dose dependent (p < 0.05) reduced uterine contractions decreased dose-dependently after of addition oxytocin. The extracts added cumulatively to the organ bath reduced contractions but they did not affect uterine contractions induced by BaCl2 except the last dose. Spasmolytic effects of the extracts were not affected by propranolol or naloxane in KCl induced contractions. CONCLUSION: Extracts diminished K(+)-induced contraction in uterus, therefore it seems that substances that decrease K(+)-induced contraction can also block voltage dependent calcium channel. The extracts did not have any effect on ß-adrenoceptors or potassium channels.