مهدی توتونچی

Oncolytic viruses, as novel and advanced tools in the field of treating various types of cancer, have played a very important role in medical developments. The term “oncolytic” refers to the ability of these viruses to destroy and damage cancer cells while preserving the surrounding healthy cells.

To conduct this study, a total of 270 initial results were collected through searching in the PubMed, Scopus, and Google Scholar databases from 2012 to 2024. The primary researcher reviewed 68 relevant articles, extracted and summarized the contents, and finally compiled the findings.

The findings from this review study demonstrate that cancer cells possess distinct characteristics that differentiate them from normal cells, including continuous growth signaling, resistance to anti-growth signaling, evasion of apoptosis, increased angiogenesis, and invasion into other body parts. Oncolytic viruses utilize these distinctive features to selectively target and infect cancer cells. Most oncolytic viruses directly eliminate host tumor cells, resulting in viral replication and induction of host antiviral responses. Moreover, these viruses can destroy cancer cells through the production of specific proteins. The cytotoxic potential of oncolytic viruses depends on viral type, genetic manipulation, optimal virus dosage for injection, natural and induced viral tropism, and cancer cell sensitivity to various forms of cell death. The mechanism driving the selective replication of oncolytic viruses in cancer cells likely relates to defects in signaling pathways specific to tumor cells. Phase III clinical trials have demonstrated significant improvements in the treatment outcomes of various cancers, including head and neck cancer, melanoma, glioblastoma, and bladder cancer, through the use of H101 (Oncorine), T-Vec, ECHO-7, and Teserpaturev (Delytact) viruses.

Oncolytic viruses are constructed from various types of viruses and are currently being evaluated in laboratory, preclinical, and clinical stages. The use of these viruses for the treatment of cancer as a new and targeted approach has been proposed, which requires further investigation and achievement of more precise mechanisms for their better performance.

The evidence about the existence and function of ovarian stem cells (OSCs) is increasing. In a previous study, we isolated human theca stem cells (hTSCs) from the ovary of a 19-year-old patient and differentiated them into oocyte-like cells (hOLCs) under induced conditions. In this study, in order to prove the repeatability of this experiment as well as the presence of these cells in the ovaries of patients with higher reproductive age and their differentiation potential in-vitro, hTSCs were isolated from the ovaries of 20 and 38-year-old patients and their differentiation potential into hOLCs has been evaluated. In this experimental interventional study, based on the instructions of the previous study, hTSCs were isolated from small antral follicles with a size of 3 to 5 mm. These cells were cultured in six-well plates with A number of 5×104 cells per well in DMEM/F12 induction medium containing FBS, human follicular fluid, glutamine and pyruvate for 40 days. Then their development process was measured by assessing the morphology, size and viability. hTSCs were successfully isolated from ovarian tissue of 20 and 38-year-old patients and cultured in DMEM/F-12 medium containing growth factors (EGF, FGF, GDNF, etc.). After 12 days, hTSCs in both patients started to differentiate into hOLCs and their morphology changed from spindle-shaped to round. The size of hOLCs increased during the differentiation period in both patients (from 20-25 µm to 50 µm). The survival of hOLCs compared to hTSCs was similar in all three patients and did not differ significantly. hTSCs can be isolated from ovaries of women of different reproductive ages and there is no difference in their differentiation pattern to hOLCs in laboratory (in-vitro) conditions.

Placentome in cattle comprises fetal cotyledon and maternal caruncle and is a vital site for feto-maternal substance exchanges. There are considerable metabolic and hormonal interactions in the placentomes that could affect the body of the fetus and the mother. Despite the importance of this tissue in reproduction of cattle, there are few studies on this tissue probably due to worries about the invasive trait of placentome collection and its implications for the future reproductive performances of the cow. In this study, we described a method of placentome collection from cows and investigated its implications for the cows’ subsequent reproductive performance. In this study, placentomes were collected from six cows within 30 minutes after parturition by using a guitar string and a hygienic protocol. Then, one dose of vitamin K and two doses of B-complex (during two days) were administered and the health status of the cows was monitored for several days. The number of inseminations until conception, the length of open days, the interval between two subsequent parturitions and the health status of the following calf were recorded for each cow. These six cows considered as placentome collected group and their data were compared to 45 cows in the same herd as control group. Cows in the control group had at most 430 days of interval between their two subsequent parturitions and underwent no manipulation. According to the results, general health of the placentome collected cows was not affected after placentomectomy. None of the reproduction related parameters including the number of insemination until conception, the length of open days, the interval between two parturitions and the health status of the calves showed significant difference between the placentome collected group compared to control. However, the interval between the first parturition and the second insemination was significantly shorter in the cows that underwent placentome collection. In this study, we described an easy and efficient method of placentome collection from alive cows and showed that this method does not have considerable implications for the cows’ reproductive performance. Therefore, by careful collection of placentomes and by having justifiable research purposes, placentome collection from the cows could be considered for future studies.

Genome editing is an efficient and accurate method in biological and medical studies. Among the wide range of genome editing techniques, clustered regularly interspaced short palindromic repeats (CRISPR) is one of the simplest and yet promising methods. The CRISPR system consists of two key components; an endonuclease enzyme called Cas9 and guide RNA (gRNA), ensuring that Cas9 enzyme cuts at the right point in the genome. Although CRISPR genome editing is one of the useful methods to modify the genome, there are still multiple challenges in the technique steps.

sgRNAs were designed and ordered accordingly on the basis of the target site and vector of interest. Cloning procedures were performed and confirmed by sequencing as a gold standard. The list of high efficient sgRNAs for the LAMP-2 gene was prepared based on the available outstanding databases by analyzing and comparing the specificity and functionality as two main characteristics.

In this study, we tried to evaluate the main challenges in the process of preparing CRISPR/Cas9 popular vectors (pX330/pX459). A list of high efficient sgRNAs for an autophagy gene is also prepared as an example for clarification of the sgRNA design procedures.

This study tried to depict problems that may be encountered during sgRNA design and plasmid preparation, followed by giving appropriate recommendations.

Sperm associated antigens (SPAGs) play an important role in the incidence of various cancers including breast, lung, liver, and bladder. SPAGs are also important in sperm functions, such as motility. However, it seems that sperm cryopreservation as one of the assisted reproductive techniques (ART), can affect the expression of these genes. This study aimed to evaluate the effect of freezing on the expression of human SPAG 5 and 9 in human spermatozoa.

In the present experimental study, twelve semen samples in terms of normozoospermic parameters were collected from individuals referred to the Royan Institute, and progressive motile sperms were isolated by density gradient centrifugation (DGC). Each sample was divided into two, non-frozen (control) and frozen groups. After rapid freezing and three-day storage in liquid nitrogen, samples were thawed in tap water and incubated for 2 hours of recovery-time in a CO2 incubator. RNA extraction in both groups was performed using TRIzol; and SPAG5 and 9 were evaluated by Real-time PCR technique.

Based on statistical analysis, the expression of SPAG5 decreased significantly in the frozen group compared to the control group (P≤0.05); In contrast, there was no significant difference in the expression of SPAG9 between the control and frozen groups.

Considering the cold shock in the future of cell, a significant reduction in SPAG5 expression in the frozen group may indicate probable influences in the derived fetus.

The retinal pigment epithelium cells (RPE) have crucial roles in the health and functionality of retina. Any damage or dysfunction of these cells can lead to severe retinopathies. Identification of signaling pathways and biological processes involved in RPE differentiation can be useful in devising more robust therapeutic approaches.

In the present study, we used the intersection of three online prediction databases and their ::union:: with one experimental database to select microRNAs gene targets. Next, by the intersect of the targeted genes with an increase in their expression in epithelial to mesenchymal transition (EMT) of RPE cells, we tried to build a microRNA-mRNA integrative network. Further, several pathway analyses tools were used to perform a more accurate and comprehensive analysis of the signaling pathways and biological processes being regulated by selected miRs in the EMT of the RPE cells.

Our study revealed that among the 3406 genes being upregulated over the course of EMT in RPE cells, adj p-value≤0.05, fold change≥1.5, 93 genes were miR-204-5p and miR 211-5p target genes. Further analysis of the obtained target gene list demonstrated that these two microRNAs are mostly involved in maintaining RPE cells from going through EMT via regulation of cell adhesion and secretion subnetworks and also MAPK and TGF-β1 signaling pathways while preserving cells from apoptosis and neuronal fates.

This study indicated that miR-204-5p and miR 211-5p are involved in protecting RPE cells from EMT and reinforce their epithelial cell identity.

There is increasing evidence that cancer cells in addition to multiple genetic mutations, also acquire epigenetic abnormalities during development, maintenance, and progression. By utilizing the reprogramming technology as a tool to introduce the ‘pressure’ to alter epigenetic regulations, we might be able to clarify the epigenetic behavior that is unique to cancer cells. So far, iPSCs have been generated from normal primary cells, but it is unclear whether human primary cancer cell can be reprogrammed. We investigated the production of the iPS cells from the pancreatic adenocarcinoma cells using defined transcription factors. We sought to reprogram patient derived xenograft from human PDAC, by introducing lentiviral mediated induction of Yamanaka Factors (OSKM) and characterized of induced cells by Alkaline Phosphatase staining, Real-Time PCR and immunostaining.  This study with research ethics code EC/93/1025 has been approved by research ethics committee at Royan Institute. Alkaline Phosphatase staining, Real-Time PCR and immunostaining showed that induction with the OSKM results in generating iPS cell line from fibroblast cells but not from PDAC PDX cells .We showed that, PDAC cells could not fully reprogrammed by the expression of 4 transcription factors. This study demonstrated that the PDAC-PDX cancer cells were distinct from PDAC induced cells with regard to their epigenetic modifier genes expression pattern, although the expression of pluripotency genes did not increased significantly in the induced PDAC cells.

Asthenozoospermia, as the most prevalent cause of male infertility, is defined as low percentage of progressively motile spermatozoa per ejaculate. It occurs in both non-syndromic and syndromic forms and later it manifests as a part of primary ciliary dyskinesia. In the last decade, with the advent of Next-generation sequencing technologies numerous genes have been introduced in the pathogenesis of different diseases. Here, we review the genes implicated in asthenozoospermia by genetic studies. Strategies employed by infertility genetics studies in original research articles extracted from PubMed database are critically reviewed. Afterwards, genes implicated in asthenozoospermia and primary ciliary dyskinesia are discussed. Until today, pathogenic variants in DNAH1, SEPT12, SLC26A8, CATSPER1, CATSPER2 and ADCY10 have been reported to cause non-syndromic asthenozoospermia. Moreover, DNAI1, DNAH5, DNAAF2, CCDC39, DYC1X1 and LRRC6 have been implicated in primary ciliary dyskinesia and syndromic asthenozoospermia. Next-generation sequencing technologies and especially whole-exome sequencing in families with multiple asthenozoospermic patients showed considerable success in introduction of genes involved in asthenozoospermia leading to a more comprehensive knowledge on genetics of infertility.

Cancer stem cells are responsible for the formation the resistance to treatment, tumor relapse, and metastasis. miRNAs play an important role in the regulation of biological processes. Therefore, the purpose of this review is to candidate miRNAs that are involved in the regulation of all three properties including stemness, metastasis, and drug resistance and find their target genes and signaling pathways by using literature learning and data mining. The present systematic review is done to identify stemness-regulating miRNAs. By using CORMINE database, metastasis and drug resistance regulating miRNAs collected. Finally, we compared these three lists of miRNAs and found common miRNAs in these three properties. ONCO.IO database and KEGG pathway have been done to obtain the interaction between miRNA-miRNA target and cancer-related signaling pathway respectively. We collected 191 stemness-regulating miRNAs from 21 excluded studies. Based on CORMINE database, 161 miRNAs and 57 miRNAs had metastasis and stemness features respectively. We obtained 7 common miRNAs that 4 of them including has-miR-34a, has-miR-23a, has-miR-30a, has-miR-100 has a significant role for targeting signaling pathways involved in cancer and their most important targets regulate many processes of cells. These data suggest that three important properties can regulate by common miRNAs. Therefore, target these miRNAs or their targets can be helpful to stop tumor growth and metastasis and may be useful biomarkers for early detection of gastric cancer.

Stem cells are believed as the premier hope to regenerate the defected tissues. In this regard, enhancing the efficient derivation of mouse embryonic stem (ES) cells, as the best model of pluripotent stem cell, facilitates capturing of the efficient derivation of human ES cells. Small molecules play a critical role in improving the efficiency of generating the pluripotent stem cells by inhibiting signaling pathways related to differentiation. This study aimed to evaluate the role of some molecular signaling pathways (e.g. JAK/STAT, MAPK/ERK, PI3K/AKT, WNT/GSK3 and TGF-β) in the cells involved in producing and maintaining ES cells. In this review study, the relevant articles to signaling pathways and embryonic stem cells were selected from PubMed database. In addition, the procedure for the efficient derivation of mouse ES cells was analyzed using small molecules under different conditions, like 2i and R2i culture. The R2i culture condition increases the efficiency of generation and maintenance of ES cell lines from different types of mouse strain. Thus, findings showed that by inhibiting the MEK and TGF-β pathways in this process, the higher frequency of cells would be maintained at ground state of pluripotency with no differentiation. To understand the molecular effects of R2i culture condition on enhancing the efficiency of generating the mouse ES cells, the assessment of key pluripotency and differentiation gene expressions as well as the epigenetic changes within the ES cell derivation process seems to be essential.

The majority of cancer treatments are invasive. Gonadal injuries cause reductions in fertility which results in lack of hope for conception in cancer patients and frustration for their partners. Fortunately، current advancements in cryopreservation and transplantation sciences regarding fertility preservation lead to cryostorage of gonads and preservation prior to the onset of chemo- and radiotherapy treatments. Accordingly in women، the main goal of ovarian cryopreservation is establishment of fertility and hormonal cycle restoration after auto-transplantation. Although the history of ovarian transplantation dates back to the 19th century، there are reports of live human births following ovarian tissue cryopreservation and transplantation since the past 100 years. Despite this success and additional research in the field of ovarian cryopreservation and transplantation، numerous questions remain unanswered. Among these questions، growth factors and hormonal changes because of their effects on follicular function appear to be more important during ovarian tissue transplantation. This review attempts to address hormones and growth factor functions with the specifics of ovarian cryopreservation and auto-transplantation.

Many changes of female reproductive system are based on the vascular activity and blood flow. In ovarian follicular development, re-anastomosis is critical and permits cells and oocytes to utilize nutrients as well as the hormones. Moreover, it plays a main role in corpus luteum formation. Female reproductive system undergoes several angiogenesis programs which are considered essential in order to reach a follicle to preovulatory stage and to set up a network between granulosa cells and blood vessels through the theca layer. Up to the first 3 days after transplantation and reanastomosis start, a large number of follicles especially advanced types, begin an early death due to their severe dependence on nutrients of blood supply. Loss of the nutritional sources of granulosa cells leads to a decrease in survival and supporting factors and thus, cell death increases. Considering the importance of this issue, the present study intends to investigate the activity of angiogenic and apoptotic factors in transplanted ovarian tissue.

Embryo cryopreservation is the process that water is removed from the cell by cryoprotectant materials, and embryos are stored at temperature below zero. This process may affect the viability and developmental potential of embryos. In this study, the effect of the vitrification cryotop method on the expression level of Oct4 and Mest developmental genes in mouse blastocysts was examined. The collected 2-cell embryos of superovulated mouse by oviduct flushing were divided into non-vitrified and vitrified groups. These embryos were cultured to the blastocyst stage directly in the non-vitrified group and in the vitrified group, these embryos were cultured to 4-8 cell embryos, vitrified with cryotop in these stages and after 2-6 months, warmed and cultured to blastocyst embryos. Quantitative expression of two developmental genes, namely Oct4 and Mest, were performed in these groups, using RNA purification and Real-time RT-PCR. Quantitative PCR analysis showed that the expression level of both genes, Oct4 and Mest, was reduced significantly in the vitrified-warmed group relative to the control group (p=0.046 and p=0.001). This study revealed that morphologically normal embryos show a reduced amount of Oct4 and Mest transcripts which indicate that the vitrification method negatively effects the expression level of these two developmental genes.

The purpose of this study was to evaluate the quantitative expressions of BAG1, BAX and BCL-2 in human embryos with different fragmentation grades as derived from assisted reproduction technology (ART). Fragmented and normal human 8-cell embryos were scored according to the degree of fragmentation with an inverted microscope and divided into four grades (grade І: no or minimal fragmentation (<5%), grade ІІ: embryos with <25% fragmentation, grade ІІІ: embryos with >25% fragmentation and grade ІV: apoptotic induced embryos with actinomycin D). In this study, TUNEL labeling was initially used to detect apoptosis, and then revers transcription polymerase chain reaction (RT-PCR) and quantitative PCR were used to define the quantitative expressions of experimental genes in human embryos with different fragmentation grades. The results of TUNEL labeling showed that embryos with higher fragmentation had a high number of apoptotic bodies. The results of RT-PCR and q-PCR analyses showed a significantly decreased amount of BAGI transcript expression from group I to group IV. The highest expression of BAX gene was observed in group II, however, the transcript of BCL-2 gene was not observed in any of the experimental groups. The effect of actinomycin D on transcript expression amounts of experimental genes in apoptotic induced embryos (group IV) compared to control embryos (group I) showed a significant decrease. mRNA expression of BAG1 gene can be used as a good marker to detect apoptosis in human embryos. However, the transcript of BCL-2 gene does not play a role in the detection of apoptosis in human embryos at the 8-cell stage.