Enhancing the persistence and memory of CAR T cells

CAR T cell therapy has revolutionized cancer treatment, but has also provided an opportunity for treating chronic viral infections such as HIV, HBV, and HCV. Despite the profound outcomes in the treatment of hematological malignancies, CAR T cell therapy for solid tumours has not been almost invariably unsuccessful in the clinic. Hostile conditions of TME, low tumour infiltration, lack of persistence, and absence of memory CAR T cell formation are the main obstacles ahead of CAR T cell therapy for solid tumours. This study aimed to improve Her2-CAR T cell persistence and TM (T memory) development. TM cells have distinct mitochondria morphology and metabolism. TM cells have larger mitochondria (fusion) and rely on OXPHOS metabolism. In order to achieve the aim of this study, we selected Mcl-1 and miR429 as genes to overexpress in CAR T cells. Recently, several studies suggested that AICD (activation-induced cell death) induced by the CD95 pathway is the one of the main causes of low CAR T cell persistence in vivo. Mcl-1 is also well characterised for its role in OXPHOS metabolism, mitochondrial energetics and mitochondrial fusion. To complement this approach, the miRNA429 was selected as a means to enhance CAR T cell function through the suppression of genes that negatively affect T cell function, TM development, and mitochondrial fusion such as TCAIM, MFF, and TET-2. The first aim of this study was to upregulate the endogenous level of Mcl-1. We tested eight small activating RNA (saRNA) targeting different regions of the Mcl-1 promoter, but none of them was able to induce Mcl-1. Further promoter analysis led to the identification and characterisation of the first antisense transcript (named mcl1-AS1) that modulate Mcl-1 expression. However, due to the late manifestation of gene regulation (at 48 - 72 hours) that was seen following mcl1-AS1 inhibition, it was not applicable for us to use this strategy to Mcl-1 expression (Chapter II). The next strategy was the controlled expression of Mcl-1 using the Tet-On system. We used several approaches to improve the Tet-On system, including gene replacement, codon-optimisation of rt-TA, using G72V-rtTA, removing cryptic splice sites within rt-TA, creating an autoregulatory Tet-On system, and manipulating regulatory elements in TCE minimal promoter. Our final optimised construct showed high inducibility and a very low background expression compared to the original construct (Chapter III). However, due to the low transfection efficiency of SB system in primary T cells and lack of artificial antigen presenting cell (aAPC) at the time for expansion of T cells, we decided to create an inducible LV system. The lack of inducibility in low doxycycline concentration and low transduction efficiency made our inducible LV system not suitable for our study. Therefore, we decided to use a constitutive system to see the effects of Mcl-1 and miR429 overexpression in CAR T cells. In order to express Mcl-1 and miR429 in a constitutive LV system, we examined the strength of four commonly used promoters, EF-1, CMV, RPBSA, and hPGK, in running short and long transcripts. EF-1 showed to be the best promoter in running short and long RNA in T cells (Chapter IV). As a result, we chose EF-1 to run the GFP-P2A-Her2CAR and hPGK to transcribe Mcl-1 or miR429. For the first time, we showed that TCAIM, MFF, and TET-2 are direct targets of miR429. Overexpression of miR429 in CAR T cells slightly increased the number of TSCM and TCM in CD4+CAR T cells, while the number of Treg and TEMRA cells was marginally decreased. Upregulating Mcl-1 in CAR T cells promoted the TSCM and TCM development in both CD4+ and CD8+ CAR T cells, whereas the frequency of Treg and TEMRA cells declined. Mcl-1 overexpression also protected CAR T cells from CD95L-induced AICD. Although our study cannot provide a mechanism for the Mcl-1 role in memory CAR T cell development, an increase in mitochondrial mass and mtDNA suggest that Mcl-1 likely enhances mitochondrial energetics and fusion (Chapter V). Lastly, we are in a SARS-CoV-2 pandemic era. When locked-down and unable to access the laboratory in early 2020, we investigated the effect of recurrent mutations on viral RNA secondary structure and host miRNA interaction. From an evolutionary point of view, mutations that arise several times independently (homoplasies) and lead to clade expansion are highly likely to increase viral fitness. The emergence of several mutations has resulted in the emergence of a G-clade responsible for 97% of cases around the globe. This clade consists of four mutations, two cause amino changes (C14408U and A23403G), while others (C241U, C3037U) are silent and are currently of unknown impact on viral fitness. Based on our bioinformatics survey, the C3037U mutation destroys the miR-197-5p binding site. Interestingly, miR-197-5p is highly expressed in SARS-CoV-2 target cells and has been reported to be upregulated in patients with cardiovascular disease. Interestingly, this miRNA also acts as defence against variety of viral infections such as HBV, HCV, HAV, Enterovirus 71, Ebola and H7N9. Further in vitro work is underway to test the significance of the C3037U mutation on miRNA inhibition in a SARS-2 pseudovirus and live virus assays

Tunability of terahertz random lasers with temperature based on superconducting materials

We theoretically demonstrate the tunabiltiy of terahertz random lasers composed of high temperature superconductorYBCO and ruby layers as active medium. The considered system is a one-dimensional disordered medium made of ruby grain and YBCO. Finite-difference time domain method is used to calculate the emission spectrum and spatial distribution of electric field at different temperatures. Our numerical results reveal that the superconductor based random lasers exhibit large temperature tunability in the terahertz domain. The emission spectrum is significantly temperature dependent, the number of lasing modes and their intensities increase with decreasing temperature. Also, we make some discussion to explain the reason for the observed tunability and the effect of temperature variation on the spatial distribution of the electric field in the disordered active medium

Planning of Sustainable Development Based on Civil Rights in Torkaman Port

Undoubtedly, discussion about stability and sustainable development is meaningless without paying attention to cities and urbanization. Cities are considered as major factors of creating instability in the world. Therefore, according to inherent complexity of cities and various dimensions of their effectiveness, recognition of major and key factors for the sake of achieving to stability of cities seems necessary. The purpose of research is to acquire relatively exact description of condition of urban sustainable development from citizens’ rights point of view. This research is practical from the viewpoint of purpose and descriptive-measurable from the viewpoint of type, which has been done by scope method. Statistical society is all citizens of Torkaman city (48736) that used urban spaces which are effective on stability and instability of city. Sample volume is 381 individuals according to Cochran formula that are selected and studied accidentally. Findings of research show that civil rights identifiers with effectiveness of 0.62 and political identifiers with effectiveness of 0.18 had the most and the least effect on sustainable development of Torkaman port city

Planning of Sustainable Development Based on Civil Rights in Torkaman Port

Undoubtedly, discussion about stability and sustainable development is meaningless without paying attention to cities and urbanization. Cities are considered as major factors of creating instability in the world. Therefore, according to inherent complexity of cities and various dimensions of their effectiveness, recognition of major and key factors for the sake of achieving to stability of cities seems necessary. The purpose of research is to acquire relatively exact description of condition of urban sustainable development from citizens’ rights point of view. This research is practical from the viewpoint of purpose and descriptive-measurable from the viewpoint of type, which has been done by scope method. Statistical society is all citizens of Torkaman city (48736) that used urban spaces which are effective on stability and instability of city. Sample volume is 381 individuals according to Cochran formula that are selected and studied accidentally. Findings of research show that civil rights identifiers with effectiveness of 0.62 and political identifiers with effectiveness of 0.18 had the most and the least effect on sustainable development of Torkaman port city

Evaluating the Effect of Family-Centered Intervention Program on Care Burden and Self-Efficacy of Hemodialysis Patient Caregivers Based on Social Cognitive Theory: A Randomized Clinical Trial Study

Background Caregivers of hemodialysis patients spend a large amount of time providing care to these patients while tolerating fatigue and stress. This study evaluated a family-centered empowerment program on the care burden and self-efficacy of hemodialysis patient caregivers based on social cognitive theory. Methods In this randomized clinical trial, 70 family caregivers of hemodialysis patients in Isfahan, Iran, were selected and randomly allocated to intervention and control groups, in 2015–2016. Two questionnaires were used to collect the family caregivers’ characteristics, care burden, and self-efficacy, and patients’ negative and positive outcomes expectancies. Data were analyzed using SPSS before, immediately after, and 2 months after the intervention. Results There was no significant difference in the mean scores of care burden, positive outcomes expectancies, negative outcomes expectancies, and self-efficacy between the two groups before the intervention. However, there were significant differences in the post-test and follow-up data analyses (P<0.05). Conclusion Given the degenerative nature of chronic kidney disease, it can be considered as a source of long-term and chronic stress for caregivers. Therefore, by implementing an empowerment program, caregiving behaviors can be improved, positive outcomes expectancies can be increased, and negative outcomes expectancies can be reduced. Keywords: Family-Centered Nursing; Illness Burdens; Family Caregivers; Hemodialysis Patien