CAR-T cell immunotherapy for ovarian cancer: hushing the silent killer.

As the most lethal gynecologic oncological indication, carcinoma of the ovary has been ranked as the 5th cause of cancer-related mortality in women, with a high percentage of the patients being diagnosed at late stages of the disease and a five-year survival of ~ 30%. Ovarian cancer patients conventionally undergo surgery for tumor removal followed by platinum- or taxane-based chemotherapy; however, a high percentage of patients experience tumor relapse. Cancer immunotherapy has been regarded as a silver lining in the treatment of patients with various immunological or oncological indications; however, mirvetuximab soravtansine (a folate receptor α-specific mAb) and bevacizumab (a VEGF-A-specific mAb) are the only immunotherapeutics approved for the treatment of ovarian cancer patients. Chimeric antigen receptor T-cell (CAR-T) therapy has achieved tremendous clinical success in the treatment of patients with certain B-cell lymphomas and leukemias, as well as multiple myeloma. In the context of solid tumors, CAR-T therapies face serious obstacles that limit their therapeutic benefit. Such hindrances include the immunosuppressive nature of solid tumors, impaired tumor infiltration, lack of qualified tumor-associated antigens, and compromised stimulation and persistence of CAR-Ts following administration. Over the past years, researchers have made arduous attempts to apply CAR-T therapy to ovarian cancer. In this review, we outline the principles of CAR-T therapy and then highlight its limitations in the context of solid tumors. Ultimately, we focus on preclinical and clinical findings achieved in CAR-T-mediated targeting of different ovarian cancer-associated target antigens.

T-cells engineered with a novel VHH-based chimeric antigen receptor against CD19 exhibit comparable tumoricidal efficacy to their FMC63-based counterparts.

Background: Chimeric antigen receptor (CAR)-T cell therapy has established itself as a potent therapeutic option for certain patients with relapsed/refractory (R/R) hematologic malignancies. To date, four CD19-redirected CAR-T cell products have been granted the United States Food and Drug Administration (FDA) approval for medical use. However, all of these products are equipped with a single-chain fragment variable (scFv) as their targeting domains. Camelid single-domain antibodies (VHH or nanobody) can also be used as alternatives to scFvs. In this study, we developed VHH-based CD19-redirected CAR-Ts, and compared them with their FMC63 scFv-based counterpart. Methods: Human primary T cells were transduced to express a second-generation 4-1BB-CD3ζ-based CAR construct whose targeting domain was based on a CD19-specific VHH. The expansion rate, cytotoxicity, and secretion of proinflammatory cytokines (IFN-γ, IL-2, and TNF-α) of the developed CAR-Ts were assessed and compared with their FMC63 scFv-based counterpart as they were co-cultured with CD19-positive (Raji and Ramos) and CD19-negative (K562) cell lines. Results: VHH-CAR-Ts showed an expansion rate comparable to that of the scFv-CAR-Ts. In terms of cytotoxicity, VHH-CAR-Ts mediated cytolytic reactions against CD19-positive cell lines, comparable to those of their scFv-based counterparts. Moreover, both VHH-CAR-Ts and scFv-CAR-Ts secreted remarkably higher and similar levels of IFN-γ, IL-2, and TNF-α upon co-cultivation with Ramos and Raji cell lines compared with while cultured alone or co-cultured with K562 cells. Conclusion: Our results demonstrated that our VHH-CAR-Ts could mediate CD19-dependent tumoricidal reactions as potently as their scFv-based counterparts. Moreover, VHHs could be applied as the targeting domains of CAR constructs to overcome the issues associated with the use of scFvs in CAR-T therapies.

Effects of polybrene and retronectin as transduction enhancers on the development and phenotypic characteristics of VHH-based CD19-redirected CAR T cells: a comparative investigation.

Chimeric antigen receptor T cells (CAR T cells) have improved the prognosis of patients with certain hematologic malignancies. However, broader clinical application of this type of therapy is dependent on production protocols. We characterized VHH-based CD19-redirected CAR T cells generated using the transduction enhancers (TEs) polybrene or retronectin. The proliferation rate of activated T cells transduced using polybrene concentrations > 6 mg/mL decreased compared with untreated group. There was a direct relationship between polybrene concentration and transduction efficacy. Moreover, we demonstrated the proliferation of retronectin-transduced T cells increased in a dose-dependent manner (4-20 µg/mL). Whereas, different retronectin concentrations did not mediate a significant increase in T cell transduction rate. Moreover, lentiviral transduction rate was also dependent on the concentration of lentiviruses. At optimized TE concentrations, multiplicity of infection (MOI) of > 10 decreased living T cell transduction rate. Additionally, we demonstrated that CAR T cell phenotype is highly affected by TE type. Naïve T cell differentiation to central memory T cell was observed in the beginning of the expansion process and effector memory T cells became the predominant subset in the second week of expansion. Importantly, retronectin increased the proliferation of CAR T cells alongside medicating higher transduction rates, resulting in more naïve and central memory T cells. We demonstrated that a higher percentage of CAR T cells were generated using retronectin (with a less differentiated phenotype) making retronectin a more effective TE than polybrene for long-term CAR T cell processing in preclinical or clinical studies.

CAR-T cell therapy in triple-negative breast cancer: Hunting the invisible devil.

Triple-negative breast cancer (TNBC) is known as the most intricate and hard-to-treat subtype of breast cancer. TNBC cells do not express the well-known estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2 (HER2) expressed by other breast cancer subtypes. This phenomenon leaves no room for novel treatment approaches including endocrine and HER2-specific antibody therapies. To date, surgery, radiotherapy, and systemic chemotherapy remain the principal therapy options for TNBC treatment. However, in numerous cases, these approaches either result in minimal clinical benefit or are nonfunctional, resulting in disease recurrence and poor prognosis. Nowadays, chimeric antigen receptor T cell (CAR-T) therapy is becoming more established as an option for the treatment of various types of hematologic malignancies. CAR-Ts are genetically engineered T lymphocytes that employ the body's immune system mechanisms to selectively recognize cancer cells expressing tumor-associated antigens (TAAs) of interest and efficiently eliminate them. However, despite the clinical triumph of CAR-T therapy in hematologic neoplasms, CAR-T therapy of solid tumors, including TNBC, has been much more challenging. In this review, we will discuss the success of CAR-T therapy in hematological neoplasms and its caveats in solid tumors, and then we summarize the potential CAR-T targetable TAAs in TNBC studied in different investigational stages.

Hybrid powdered activated carbon-activated sludge biofilm formation to mitigate biofouling in dynamic membrane bioreactor for wastewater treatment.

Membrane costs and biofouling limit applications of membrane bioreactors (MBRs) for wastewater treatment. Here, powdered activated carbon (PAC) utilization in the formation and performance of a self-forming dynamic membrane consisting of activated sludge and PAC during hybrid wastewater treatment process was studied. Short-term agitation helped (non)biological particles to quickly uniformly settle on mesh filter, forming more uniform PAC-containing dynamic membranes (PAC-DMs). PAC adsorbed adhesive materials, resulting in an increase in average floc size and DM permeability while decreasing biofouling. The most efficient PAC concentration was 4 g L-1 considering techno-economics, i.e. the highest effluent quality (turbidity of 19.89 NTU) and the lowest biofouling (transmembrane pressure rise of 2.89 mbar). Short-term performance of hybrid PAC-DM bioreactor (PAC-DMBR) showed stability in effluent quality improvement including 92%, 95%, 83%, 84% and 98% reductions in turbidity, chemical oxygen demand, total dissolved solids, total nitrogen, and total phosphorous, respectively. Accordingly, adopting hybrid PAC-DMBR has potential to alleviate biofouling and capital cost.

A split-face comparison of fractional erbium: YAG laser plus Kligman's formula vs. Kligman's formula monotherapy for facial melasma.

Melasma is a chronic hyperpigmentation disorder. Although several treatment methods are used for patients, it remains a challenging problem for physicians. Erbium-YAG laser is one of the laser treatment methods that proved its efficacy in melasma treatment. We aimed to compare the efficacy of combining the fractional Er: YAG laser and Kligman's formula with Kligman's formula alone in the treatment of melasma. Twenty female patients with bilateral melasma were randomly treated in a split-face controlled manner with a fractional Er: YAG laser followed by Kligman's formula on one side and Kligman's formula on the other side. All patients received three laser sessions with four-week intervals. The efficacy of treatments was determined through photographs, Visioface, and Melasma Area Severity Index (MASI) score, all performed at baseline and three months after the end of laser sessions. Fractional Er: YAG laser and Kligman's formula showed a significant decrease in MASI score than Kligman's formula alone (P = 0.005). There was a significant decrease in cheek Visioface score on Er: YAG laser and Kligman's formula vs Kligman's formula (P = 0.02). However, the Patient Global Assessment Scale of both sides was not statistically significant (P = 0.23). The combination of Fractional Er: YAG laser with Kligman's formula is an effective treatment method for melasma.

A Patient with Trisomy 4p and Monosomy 10q

Translocations are the most common structural abnormality in the human genome. Carriers of balanced chromosome rearrangements exhibit increased risk of abortion or a chromosomally-unbalanced child. The present study was carried out in 2017 at the Iranian Blood Transfusion Research Center. This study reported a rare chromosomal disorder with 4p duplication and 10q distal deletion syndrome which is associated with various complications at birth. Defects included the following characteristics: dysmorphic facial characteristic, hand or foot anomalies, growth retardation, developmental delay, strabismus, heart defects and renal anomalies. Cytogenetic analysis and array CGH were performed and, for the first time, we reported a patient with trisomy 4p16.3p12 and monosomy 10q26.3. The patient was found to have: arr 4p16.3p12 (37,152-45,490,207) x3, 10q26.3 (134,872,562-135,434,149) x1 genomic imbalances.

Electrospun Poly-ε-Caprolactone (PCL)/Dicalcium Phosphate Dihydrate (DCPD) Composite Scaffold for Tissue Engineering Application.

Recently electrospun scaffolds show excellent response in cell adhesion, growth, and tissue healing in comparison with other techniques. So in this study, PCL and PCL/DCPD scaffolds were designed and prepared with electrospinning. The electrospun scaffolds were characterized by scanning electron microscope with X-ray elemental analysis, atomic force microcopy, differential scanning calorimetry, and contact angle analysis for optimizing the effective parameters. Fiber formation with uniform diameter and bead-free structure was obtained. Scaffold surface roughness increased from 100 nm for PCL to 440 nm for PCL/DCPD. DSC analysis showed the effects of DCPD on thermal stability of composite scaffold and the results of contact angle evaluation indicate improved hydrophilicity and ability of water absorption of PCL/DCPD composite fibers as compared to PCL fibers. MTT assay indicated lack of toxicity for human gingival fibroblast (HGF) cells after cell seeding on scaffold. Also, the composite scaffold can improve cell viability by helping their growth on its surface. So it can be concluded that by engineering the electrospinning parameters we can fabricate a PCL/DCPD composite scaffold for tissue engineering applications.

Autophagy-Modulated Human Bone Marrow-Derived Mesenchymal Stem Cells Accelerate Liver Restoration in Mouse Models of Acute Liver Failure.

BACKGROUND: Mesenchymal stem cells (MSCs) have been recently received increasing attention for cell-based therapy, especially in regenerative medicine. However, the low survival rate of these cells restricts their therapeutic applications. It is hypothesized that autophagy might play an important role in cellular homeostasis and survival. This study aims to investigate the regenerative potentials of autophagy-modulated MSCs for the treatment of acute liver failure (ALF) in mice. METHODS: ALF was induced in mice by intraperitoneal injection of 1.5 ml/kg carbon tetrachloride. Mice were intravenously infused with MSCs, which were suppressed in their autophagy pathway. Blood and liver samples were collected at different intervals (24, 48 and 72 h) after the transplantation of MSCs. Both the liver enzymes and tissue necrosis levels were evaluated using biochemical and histopathological assessments. The survival rate of the transplanted mice was also recorded during one week. RESULTS: Biochemical and pathological results indicated that 1.5 ml/kg carbon tetrachloride induces ALF in mice. A significant reduction of liver enzymes and necrosis score were observed in autophagy-modulated MSC-transplanted mice compared to sham (with no cell therapy) after 24 h. After 72 h, liver enzymes reached their normal levels in mice transplanted with autophagy-suppressed MSCs. Interestingly, normal histology without necrosis was also observed. CONCLUSION: Autophagy suppression in MSCs ameliorates their liver regeneration potentials due to paracrine effects and might be suggested as a new strategy for the improvement of cell therapy in ALF.

Sticker reminders improve thromboprophylaxis appropriateness in hospitalized patients.

BACKGROUND: Venous thromboembolism (VTE) is a major health problem. Even though effective thromboprophylaxis measures exist to prevent VTE, close adherence to guidelines is missing. We assessed the effects of pasting VTE prophylaxis sticker reminders, on the appropriateness of thromboprophylaxis and prophylaxis underutilization. METHODS: Thromboprophylaxis reception was sought prospectively in two time points before and two time points after pasting sticker reminders in hospitalized patients of Masih Daneshvari Medical Center, Tehran, Iran. Thromboprophylaxis reception appropriateness was evaluated by the eighth American College of Chest Physicians (ACCP) guidelines on antithrombotic and thrombolytic therapy. Co-morbidities and conditions considered to affect the risk of venous thromboembolism were also recorded. RESULTS: Prophylaxis reception and appropriateness were studied in 298 patients before and 306 patients after the intervention. Based on the ACCP guidelines, overall thromboprophylaxis appropriateness was improved after the intervention (70.4% before, and 78.1% after the intervention, P=0.03). Prophylaxis underutilization, and prophylaxis initiation delay in those who needed thromboprophylaxis, were also reduced (P=0.03, and P=0.011 respectively). The intervention did not result in an increased rate of overprophylaxis (P=0.45). CONCLUSION: Sticker reminders could be safely and effectively incorporated into strategies to improve VTE prophylaxis and prophylaxis appropriateness, particularly in healthcare settings where electronic alert systems are not available.

Cytogenetic investigation in Iranian patients suspected with Fanconi anemia.

We present our study on 318 patients suspected with Fanconi anemia (FA) referred to The Iranian Blood Transfusion Organization during the period of 4 years. Mitomycin C (MMC) was used as a DNA cross-linker to study chromosomal breakage. In total 61 positive cases were diagnosed cytogenetically. The ratio of women being affected was slightly higher than men. Comparison of several hematologic and clinical parameters in FA (MMC positive) and non-FA (MMC negative) patients showed no clinically significant differences. This study also indicates that this sort of test is very useful and essential for accurate diagnosis of patients with FA with or without congenital anomalies.