Novelty in improvement of CAR T cell-based immunotherapy with the aid of CRISPR system

Abstract Introduction Chimeric Antigen Receptor (CAR) T cells have tremendous potentials for cancer treatment; however, various challenges impede their universal use. These restrictions include the poor function of T cells in tumor microenvironments, the shortage of tumor-specific antigens and, finally, the high cost and time-consuming process, as well as the poor scalability of the method. Creative gene-editing tools have addressed each of these limitations and introduced next generation products for cell therapy. The clustered regularly interspaced short palindromic repeats-associated endonuclease 9 (CRISPR/Cas9) system has triggered a revolution in biology fields, as it has a great capacity for genetic manipulation. Method In this review, we considered the latest development of CRISPR/Cas9 methods for the chimeric antigen receptor T cell (CAR T)-based immunotherapy. Results The ability of the CRISPR/Cas9 system to generate the universal CAR T cells and also potent T cells that are persistent against exhaustion and inhibition was explored. Conclusion: We explained CRISPR delivery methods, as well as addressing safety concerns related to the use of the CRISPR/Cas9 system and their potential solutions.

Novelty in improvement of CAR T cell-based immunotherapy with the aid of CRISPR system.

INTRODUCTION: Chimeric Antigen Receptor (CAR) T cells have tremendous potentials for cancer treatment; however, various challenges impede their universal use. These restrictions include the poor function of T cells in tumor microenvironments, the shortage of tumor-specific antigens and, finally, the high cost and time-consuming process, as well as the poor scalability of the method. Creative gene-editing tools have addressed each of these limitations and introduced next generation products for cell therapy. The clustered regularly interspaced short palindromic repeats-associated endonuclease 9 (CRISPR/Cas9) system has triggered a revolution in biology fields, as it has a great capacity for genetic manipulation. METHOD: In this review, we considered the latest development of CRISPR/Cas9 methods for the chimeric antigen receptor T cell (CAR T)-based immunotherapy. RESULTS: The ability of the CRISPR/Cas9 system to generate the universal CAR T cells and also potent T cells that are persistent against exhaustion and inhibition was explored. CONCLUSION: We explained CRISPR delivery methods, as well as addressing safety concerns related to the use of the CRISPR/Cas9 system and their potential solutions.

Cell-Based Therapy Approaches in Treatment of Non-obstructive Azoospermia.

The rate of infertility has globally increased in recent years for a variety of reasons. One of the main causes of infertility in men is azoospermia that is defined by the absence of sperm in the ejaculate and classified into two categories: obstructive azoospermia and non-obstructive azoospermia. In non-obstructive azoospermia, genital ducts are not obstructed, but the testicles do not produce sperm at all, due to various reasons. Non-obstructive azoospermia in most cases has no therapeutic options other than assisted reproductive techniques, which in most cases require sperm donors. Here we discuss cell-based therapy approaches to restore fertility in men with non-obstructive azoospermia including cell-based therapies of non-obstructive azoospermia using regenerative medicine and cell-based therapies of non-obstructive azoospermia by paracrine and anti-inflammatory pathway, technical and ethical challenges for using different cell sources and alternative options will be described, and then the more effectual approaches will be mentioned as future trends.

Comparative Evaluation of Conventional and BACTEC Methods for Detection of Bacterial Infection.

BACKGROUND: Infectious diseases are a leading cause of morbidity and mortality in developing countries. The aim of this study was to compare the results of blood culture employing the conventional and BACTEC methods for detection of bacterial infection in Taleghani Hospital, Tehran. MATERIALS AND METHODS: This was a descriptive study carried out for 3 months (March 2014-May 2014) on 272 inpatients. Their blood culture results were analyzed using the two methods (BACTEC and conventional).The results were analyzed using descriptive statistics (frequency, mean and standard deviation) and inferential tests (cross tab) via SPSS version 17 software. RESULTS: The results of 177 cases (94.1%) out of 271 studied subjects were true positive, 11 (5.8%) were false negative, 2 cases (3.15%) were false positive, and 11 cases (6.48%) were true negative. The sensitivity and specificity of the BACTEC test were 84.6 and 94.1, respectively, and the rate of positive blood cultures employing BACTEC method was equal to 100% (22.22) while in the conventional method, positive results were equal to 59.09% (22.13). CONCLUSION: Both BACTEC and conventional methods have high validity. In order to evaluate the results of blood culture and infection control, experts can use either of these methods to study the results of bacterial blood culture.

Lack of association between interleukin 28B gene polymorphisms (rs8099917G/T, rs12979860 C/T) and susceptibility to chronic hepatitis C virus infection, Tehran, Iran.

AIM: Chronic Hepatitis C infection is a critical health problem worldwide, which caused by hepatitis C virus (HCV). Interleukin 28B (IL28B) is a determinant factor in disease progression and also susceptibility to chronic HCV infection. BACKGROUND: The most significant aim of this study is to analyze the association between IL28B gene polymorphisms with susceptibility to chronic HCV infection in Iranian population. METHODS: This study follows a case- control study design, in which, 288 patients with chronic hepatitis C and 250 healthy individuals were genotyped for IL28B polymorphisms (rs12979860 C/T and rs8099917 G/T). Studied population collected from Taleghani Haospital, Tehran. Genotyping of IL28B gene polymorphisms were performed using PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) method. 10 percent of the studied population was sequenced to validate the results. RESULTS: rs8099917 G/T and rs12979860 C/T were differently distributed in hepatitis C patients and healthy controls in the female gender. TT, TG and GG genotypes distribution in the female gender were 56.7%, 39.8% and 4.5% in cases and 67%, 31.6% and 1.4% in controls (p=0.54). Also CC, CT and TT genotypes distribution were 31.8%, 61.4% and 6.8% in cases and 51.7%, 44.9% and 3.4% in controls (p=0.2). However, there was no significant difference in the allelic frequency and genotype distribution of rs12979860 C/T and rs8099917 T/G in both HCV patients with genotype 1a and 3a. CONCLUSION: It seems that rs8099917 G/T polymorphism plays a significant role in susceptibility to chronic HCV infection in Iranian population. On the other hand, no association was found between rs12979860 C/T polymorphisms and chronic hepatitis C.

Tuberculosis in Solid Organ Transplantation.

Active tuberculosis (TB) is one of the major opportunistic infections that may occur in solid organ transplant recipients. Diagnosis and treatment of latent and active TB are challenging in this population due to lack of randomized clinical trials. In this review, we discuss the clinical manifestations of TB, diagnosis of latent TB and treatment of both latent and active TB.

Effects of Pharmacist Intervention on the Utilization of Vancomycin in a Teaching Hospital.

In order to investigate the effect of pharmacist intervention on vancomycin use, this study was performed on all patients receiving vancomycin in the intensive care unit (ICU) and hematology-oncology ward of Taleghani Educational Hospital in Tehran, Iran. Vancomycin use was assessed during a pre- and post-intervention period in accordance with the Center of Disease Control and prevention (CDC) and Infectious Diseases Society of America (IDSA) guidelines. Following the intervention, there was a significant change in appropriate initiation of vancomycin (P = 0.009) and no significant improvement was observed in adequate dosage and the duration of therapy (P = 0.15 and P = 0.54 respectively); however, informing the physician resulted in discontinuation of the drug in 50% of inappropriate cases and vancomycin dosage was adjustedin 31% of cases. Temperature charts, culture results and pre-treatment CBC tests changed significantly (P = 0.02, P = 0.009 and P = 0.04 respectively). The rate of infusion related adverse drug reactions did not decrease significantly (P = 0.06); yet in 100% of patients, these reactions were resolved after notifying the nursing team. After pharmacist intervention,vancomycin use improved in some aspects. A significant improvement in appropriate initiation of therapy was observed; however, treatments continued despite negative cultures. It is necessary to optimize the use of vancomycin by performing more educational interventions.

Prevalence of atherosclerosis in patients with inactive rheumatoid arthritis.

Patients with rheumatoid arthritis (RA) exhibit higher cardiovascular morbidity and mortality than the general population. The common carotid artery intima-media thickness (CCA-IMT) measured by ultrasound is a validated surrogate marker of atherosclerosis. We studied the prevalence of subclinical atherosclerosis in patients with inactive RA. CCA-IMT was measured at the level of the carotid bifurcation in 35 patients with RA and 35 age- and sex-matched controls. CCA-IMT measurements more than two standard deviations above the mean measurement of the control group were defined as abnormal. Patients and controls with known cardiovascular risk factors were excluded from the study. The following data were taken into consideration for each patient: age, sex, duration of RA, and medication received by the patient. These variables were adjusted in data analysis. The patients were required to have had normal C-reactive protein levels for at least 1 year and no clinical signs or symptoms of active disease. An independent t test and chi-squared test were used for statistical analysis. Nine patients with RA (25.7 %) and two controls (5.7 %) had an abnormal IMT. Patients showed more carotid atherosclerotic plaques and a higher mean CCA-IMT than did controls (48.6 vs. 14.3 % and 0.705 ± 0.140 vs. 0.580 ± 0.125, respectively). An abnormal CCA-IMT was more prevalent in patients >50 years of age with >5 years of involvement (P = 0.001). These results support the theory that chronic inflammation in patients with inactive RA is associated with atherosclerosis and suggest the use of carotid ultrasonography as a useful tool with which to establish the risk of cardiovascular disease.

Stimulant Toxicity in Children: A Retrospective Study on 147 Patients.

OBJECTIVE: To evaluate the signs/symptoms of different stimulant toxicities in children to determine differences among them. DESIGN: Observational, retrospective, single-center case-series. SETTING: The only referral hospital for pediatric poisoned patients in Tehran, Iran, covering 12.5 million permanent and 6.5 million temporary residents. PATIENTS: All children between 2007 and 2012 were evaluated. Their clinical findings, laboratory tests, electrocardiogram, and outcome were evaluated. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: A total of 147 patients were enrolled (115, 24, three, and five with methamphetamine, methylphenidate, ecstasy, and unknown stimulant toxicities, respectively). Median (interquartile range) age of the methamphetamine- intoxicated children (16 mo [22-42 mo]) was significantly less than those with methylphenidate toxicity (66 mo [33-105 mo]). Almost 79% of the patients had ingested the stimulant, whereas 9% had passively been exposed to the methamphetamine smoke. The frequency of stimulant toxicity had significantly increased during the 5-year period of the study. Restlessness, mydriasis, stereotypic movements, and talkativeness were the most common signs and symptoms. Whereas bruxism only seen in 66.7% (95% CI, 21-94%) of ecstasy users, the prevalence of restlessness, sweating, and tremor was also more in this group of children. On the other hand, mydriasis and stereotypic movements were more common in crystal-meth patients by 76.5% (95% CI, 68-83%) and 53% (95% CI, 44-62%), respectively. A comparison between different routes of crystal-meth intoxication showed that flushing was more common in those who had passively/actively smoked/inhaled methamphetamine (odds ratio, 6.3 [95% CI, 1.5-26]). Palpitation was more seen in methylphenidate toxicity by 12.5% (95% CI, 4-31%). Restlessness was more detected in toddlers, whereas talkativeness and ataxia were more common in older children. Nineteen children (21%) had prolonged QTc according to normal values in different ages. None of them died. CONCLUSIONS: More studies are warranted to evaluate the frequency and outcome of this poisoning in children. Educational preventive programs are also recommended.

Simplified acute physiology score II/acute physiology and chronic health evaluation II and prediction of the mortality and later development of complications in poisoned patients admitted to intensive care unit.

We aimed to determine the acute physiology and chronic health evaluation (APACHE) II and simplified acute physiology score (SAPS) II in poisoned patients admitted to the poisoning ICU and compare them to see which is a more sensitive and specific system for prognostication of the mortality and complications in these patients. Between February 2013 and July 2013, all patients referring to our centre with any poisoning mandating ICU admission were prospectively included. On ICU arrival, a questionnaire containing the demographic data, parameters of the APACHE II and SAPS II scores, the sum of the scores, complications during the stay and the patients' final outcome (compete recovery versus death) was filled for every single patient. A total of 195 patients were evaluated. Forty-two patients (21.5%) died. Mean SAPS and APACHE scores were 41 ± 16 and 15 ± 6, respectively. Mean SAPS and APACHE scores were significantly different between the survivors and non-survivors. Both scores could successfully prognosticate the development of the complications (p = 0.07 and 0.013, respectively). APACHE II was a better score in prediction of both mortality and later complications in the setting of poisoning ICU. APACHE >22 has a good specificity in determining the mortality and development of further complications in poisoned patients admitted to the medical toxicology ICUs. SAPS II score >59 and >43 can predict the risk of mortality and later complications in these patients, as well.

Myiasis in an 89-year-old man with non-hodgkin lymphoma.

Myiasis is due to the opportunistic dipterous larvae feeding on viable or necrotic tissues of the host occurring throughout the world. We report a case of oral myiasis in an immune-compromised patient suffering from non-Hodgkin lymphoma. We would like to emphasize that such ectoparasitic infections may happen in immunocompromised patients and oral hygiene should be evaluated in all of these patients.