Phase 1 study of CAR-37 T cells in patients with relapsed or refractory CD37+ lymphoid malignancies.

We report on the first-in-human clinical trial using chimeric antigen receptor (CAR) T-cells targeting CD37, an antigen highly expressed in B- and T-cell malignancies (clinicaltrials.gov NCT04136275). Five patients with relapsed or refractory CD37+ lymphoid malignancies were enrolled and infused with autologous CAR-37 T-cells. CAR-37 T-cells expanded in the peripheral blood of all patients and, at peak, comprised >94% of the total lymphocytes in 4/5 patients. Tumor responses were observed in 4/5 patients, with 3 complete responses, 1 mixed response, and 1 patient whose disease progressed rapidly and with relative loss of CD37 expression. Three patients experienced prolonged and severe pancytopenia, and in two of these patients, efforts to ablate CAR-37 T-cells (which were engineered to co-express truncated EGFR) with cetuximab, were unsuccessful. Hematopoiesis was restored in these two patients following allogeneic hematopoietic stem cell transplantation. No other severe, non-hematopoietic toxicities occurred. We investigated the mechanisms of profound pancytopenia and did not observe activation of CAR-37 T-cells in response to hematopoietic stem cells in vitro or hematotoxicity in humanized models. Patients with pancytopenia had sustained high levels of IL-18, with low levels of IL-18 binding protein in their peripheral blood. IL-18 levels were significantly higher in CAR-37-treated patients relative to both cytopenic and non-cytopenic cohorts of CAR-19-treated cohorts of patients. In conclusion, CAR-37 T-cells exhibited anti-tumor activity, with significant CAR expansion and cytokine production. CAR-37 T-cells may be an effective therapy in hematologic malignancies as a bridge to hematopoietic stem cell transplant.

Optimization of a flow cytometry test for routine monitoring of B cell maturation antigen targeted CAR in peripheral blood.

Chimeric antigen receptor (CAR) modified T cell therapies targeting BCMA have displayed impressive activity in the treatment of multiple myeloma. There are currently two FDA licensed products, ciltacabtagene autoleucel and idecabtagene vicleucel, for treating relapsed and refractory disease. Although correlative analyses performed by product manufacturers have been reported in clinical trials, there are limited options for reliable BCMA CAR T detection assays for physicians and researchers looking to explore it as a biomarker for clinical outcome. Given the known association of CAR T cell expansion kinetics with toxicity and response, being able to quantify BCMA CAR T cells routinely and accurately in the blood of patients can serve as a valuable asset. Here, we optimized an accurate and sensitive flow cytometry test using a PE-conjugated soluble BCMA protein, with a lower limit of quantitation of 0.19% of CD3+ T cells, suitable for use as a routine assay for monitoring the frequency of BCMA CAR T cells in the blood of patients receiving either ciltacabtagene autoleucel or idecabtagene vicleucel.

Anti-TACI single and dual-targeting CAR T cells overcome BCMA antigen loss in multiple myeloma.

Chimeric Antigen Receptor (CAR) T cells directed to B cell maturation antigen (BCMA) mediate profound responses in patients with multiple myeloma, but most patients do not achieve long-term complete remissions. In addition, recent evidence suggests that high-affinity binding to BCMA can result in on-target, off-tumor activity in the basal ganglia and can lead to fatal Parkinsonian-like disease. Here we develop CAR T cells against multiple myeloma using a binder to targeting transmembrane activator and CAML interactor (TACI) in mono and dual-specific formats with anti-BCMA. These CARs have robust, antigen-specific activity in vitro and in vivo. We also show that TACI RNA expression is limited in the basal ganglia, which may circumvent some of the toxicities recently reported with BCMA CARs. Thus, single-targeting TACI CARs may have a safer toxicity profile, whereas dual-specific BCMA-TACI CAR T cells have potential to avoid the antigen escape that can occur with single-antigen targeting.

Immunogenetic Metabolomics Reveals Key Enzymes That Modulate CAR T-cell Metabolism and Function.

Immune evasion is a critical step of cancer progression that remains a major obstacle for current T cell-based immunotherapies. Hence, we investigated whether it is possible to genetically reprogram T cells to exploit a common tumor-intrinsic evasion mechanism whereby cancer cells suppress T-cell function by generating a metabolically unfavorable tumor microenvironment (TME). In an in silico screen, we identified ADA and PDK1 as metabolic regulators. We then showed that overexpression (OE) of these genes enhanced the cytolysis of CD19-specific chimeric antigen receptor (CAR) T cells against cognate leukemia cells, and conversely, ADA or PDK1 deficiency dampened this effect. ADA-OE in CAR T cells improved cancer cytolysis under high concentrations of adenosine, the ADA substrate, and an immunosuppressive metabolite in the TME. High-throughput transcriptomics and metabolomics analysis of these CAR T cells revealed alterations of global gene expression and metabolic signatures in both ADA- and PDK1-engineered CAR T cells. Functional and immunologic analyses demonstrated that ADA-OE increased proliferation and decreased exhaustion in CD19-specific and HER2-specific CAR T cells. ADA-OE improved tumor infiltration and clearance by HER2-specific CAR T cells in an in vivo colorectal cancer model. Collectively, these data unveil systematic knowledge of metabolic reprogramming directly in CAR T cells and reveal potential targets for improving CAR T-cell therapy.

Immunogenetic metabolomics revealed key enzymes that modulate CAR-T metabolism and function.

Immune evasion is a critical step of cancer progression that remains a major obstacle for current T cell-based immunotherapies. Hence, we seek to genetically reprogram T cells to exploit a common tumor-intrinsic evasion mechanism, whereby cancer cells suppress T cell function by generating a metabolically unfavorable tumor microenvironment (TME). Specifically, we use an in silico screen to identify ADA and PDK1 as metabolic regulators, in which gene overexpression (OE) enhances the cytolysis of CD19-specific CD8 CAR-T cells against cognate leukemia cells, and conversely, ADA or PDK1 deficiency dampens such effect. ADA -OE in CAR-T cells improves cancer cytolysis under high concentrations of adenosine, the ADA substrate and an immunosuppressive metabolite in the TME. High-throughput transcriptomics and metabolomics in these CAR-Ts reveal alterations of global gene expression and metabolic signatures in both ADA- and PDK1- engineered CAR-T cells. Functional and immunological analyses demonstrate that ADA -OE increases proliferation and decreases exhaustion in α-CD19 and α-HER2 CAR-T cells. ADA-OE improves tumor infiltration and clearance by α-HER2 CAR-T cells in an in vivo colorectal cancer model. Collectively, these data unveil systematic knowledge of metabolic reprogramming directly in CAR-T cells, and reveal potential targets for improving CAR-T based cell therapy. Synopsis: The authors identify the adenosine deaminase gene (ADA) as a regulatory gene that reprograms T cell metabolism. ADA-overexpression (OE) in α-CD19 and α-HER2 CAR-T cells increases proliferation, cytotoxicity, memory, and decreases exhaustion, and ADA-OE α-HER2 CAR-T cells have enhanced clearance of HT29 human colorectal cancer tumors in vivo .

Superhydrophobic polypropylene sorbent derived from discarded face masks: A highly efficient adsorbent for oil spill sorbent.

Globally, an estimated 130 billion face masks are used and disposed of every month. Thus, recycling or upcycling discarded face masks has attracted significant attention due to economic benefits and environmental concerns. To reduce the amount of used face masks going to waste, this study features a superhydrophobic face mask prepared by simple chemical modification with environmentally preferable alkane solvents (n-hexane, n-heptane, and n-decane), that is effective as a sorbent for oil spill cleanup. All alkanes examined increased the surface roughness of the face masks and improved face mask hydrophobicity. The heptane treated face mask (at 90 °C for 1 h), can adsorbed Arabian light crude oil up to 21 times of their weight on the water surface. In addition, chloroform, toluene, gasoline, and diesel were adsorbed 18, 13, 8 and 16 times, respectively. More importantly, heptane has a high recycling efficiency as a treatment solvent and is reusable for at least 10 cycles of mask surface treatment. Consequently, this inexpensive and easily fabricated material is a promising development in waste face mask (WFM) upcycling.

Bias-corrected Hp(10)-to-Organ-Absorbed Dose Conversion Coefficients for the Epidemiological Study of Korean Radiation Workers.

Background: The effects of radiation on the health of radiation workers who are constantly susceptible to occupational exposure must be assessed based on an accurate and reliable reconstruction of organ-absorbed doses that can be calculated using personal dosimeter readings measured as Hp(10) and dose conversion coefficients. However, the data used in dose reconstruction contain significant biases arising from a lack of reality and could result in an inaccurate measure of organ-absorbed doses. Therefore, this study quantified the biases involved in organ dose reconstruction and calculated the bias-corrected Hp(10)-to-organ-absorbed dose coefficients for the use in epidemiological studies of Korean radiation workers. Materials and Methods: Two major biases were considered: (1) the bias in Hp(10) arising from the difference between the dosimeter calibration geometry and the actual exposure geometry and (2) the bias in air kerma-to-Hp(10) conversion coefficients resulting from geometric differences between the human body and slab phantom. The biases were quantified by implementing personal dosimeters on the slab and human phantoms coupled with Monte Carlo method and considered to calculate the bias-corrected Hp(10)-to-organ-absorbed dose conversion coefficients. Results and Discussion: The bias in Hp(10) was significant for large incident angles and low energies (e.g., 0.32 for right lateral at 218 keV), whereas the bias in dose coefficients was significant for the posterior-anterior (PA) geometry only (e.g., 0.79 at 218 keV). The bias-corrected Hp(10)-to-organ-absorbed dose conversion coefficients derived in this study were up to 3.09-fold greater than those from ICRP publications without considering the biases. Conclusion: The obtained results will aid future studies in assessing the health effects of occupational exposure of Korean radiation workers. The bias-corrected dose coefficients of this study can be used to calculate organ doses for Korean radiation workers based personal dose records.

Myositis ossificans causing ulnar neuropathy: a case report.

Myositis ossificans (MO) can compress peripheral nerves and cause neuropathy. We herein describe a patient with ulnar neuropathy caused by MO at the medial elbow. A 28-year-old man with a drowsy mentality and multiple organ damage following a traffic accident was admitted to our hospital. After 3 weeks of postoperative care, the patient's mental status recovered. However, he complained of severe sharp pain in his left medial forearm and fourth and fifth fingers. He exhibited weak fifth finger abduction and wrist adduction. Severe elbow joint pain was elicited during range-of-motion testing of his left elbow. Ultrasound also showed an edematous, enlarged, hypoechoic ulnar nerve lying above the MO, and the MO outwardly displaced the ulnar nerve. Elbow radiographic examination, computed tomography, and magnetic resonance imaging revealed MO development and compression of the left ulnar nerve. The patient underwent surgery; the following day, his left medial forearm pain completely disappeared with slight improvement in the motor weakness of fifth finger abduction. Ultrasound is a useful tool to easily evaluate the presence of MO and compression of peripheral nerves caused by MO.

Dorsal Epidural Gas after Lumbar Microdiskectomy Treated with CT-guided Needle Aspiration.

To present a case of unusual dorsal epidural gas (EG) accumulation after a simple lumbar microdiskectomy (MD), treated with computed tomography (CT)-guided needle aspiration. A 78-year-old woman underwent simple lumbar MD at the L3-4 level. One week after the operation, the patient complained of severe back pain radiating to the right thigh. Follow-up magnetic resonance imaging (MRI) and CT revealed huge EG formation at the dorsal L3-4 epidural space. Conservative treatment did not resolve the patient's pain. We performed CT-guided needle aspiration after 1 week of conservative treatment. The patient's pain fully resolved after aspiration, but it recurred 1 week later. Follow-up MRI and CT revealed re-accumulation of the dorsal EG at the L3-4 level. CT-guided needle aspiration was repeated, again leading to full pain resolution. Follow-up CT 6 months after the second aspiration showed no recurrent dorsal EG. The patient has been symptom-free for 1 year since the second aspiration. CT-guided needle aspiration is a safe and effective alternative to re-operation in the context of dorsal EG formation after MD.

Chest Computed Tomography Findings in Asymptomatic Patients with COVID-19.

BACKGROUND: Little is known about the damage to the respiratory system in asymptomatic patients with coronavirus disease (COVID-19). OBJECTIVE: Herein, we evaluate the findings of chest computed tomography (CT) and radiography in patients with COVID-19 who were asymptomatic. METHODS: We retrospectively investigated patients with a confirmed diagnosis of COVID-19 but who did not show any symptoms. Among the 139 patients with COVID-19 who were hospitalized in Yeungnam University Hopistal in Daegu, South Korea, 10 (7.2%) were asymptomatic. Their chest CT and radiographic findings were analyzed. RESULTS: In the results, all patients (100%) had ground-glass opacity (GGO) on chest CT. Further, the GGO lesions were predominantly distributed peripherally and posteriorly in all patients. In 9 (90%) patients, the GGO lesions were combined with reticular opacity. Air bronchogram due to bronchiolectasis surrounded by GGO was observed in 8 patients (80%). Additionally, the lung lesions were dominant on the right side in all patients. CONCLUSIONS: In conclusion, considering our results that the lung is affected in asymptomatic patients, it will be necessary to extend the indications of COVID-19 testing for effective management of COVID-19 during the pandemic.

Performance evaluation of a Compton SPECT imager for determining the position and distribution of 225Ac in targeted alpha therapy: A Monte Carlo simulation based phantom study.

In this study, the performance of a Compton Single Photon Emission Computed Tomography (SPECT) imager when in vivo monitoring the position and distribution of 225Ac radionuclide in targeted alpha therapy (TAT) was evaluated. When 225Ac radionuclide, which emits various γ-rays (218 and 440 keV), is used in TAT, both the photoelectric and Compton scattering events can be used for image reconstruction. Moreover, all information pertaining to the various γ-rays of the 225Ac radionuclide can be individually or simultaneously utilized in the reconstructed image. Three types of simulation phantoms and a quantitative evaluation method were used to compare the performance of the Compton SPECT imager to that of conventional SPECT imaging, which uses only photoelectric events, and the results demonstrated that the Compton SPECT imager exhibited a higher performance as the effective count for the image reconstruction was higher. To verify the accuracy of the position and distribution of the 225Ac radionuclide that had been inserted into the phantom, reconstructed images of the various γ-rays were combined with cross-sectional images of the human phantom and all combined images were found to match the predetermined simulation conditions. In conclusion, the simulation results demonstrated the feasibility of the in vivo monitoring of the position and distribution of 225Ac radionuclide using the γ-rays in TAT.

Numerical Study on Alternating Current Breakdown Mechanism Between Sphere-Sphere Electrodes in Transformer Oil-Based Magnetic Nanofluids.

A numerical simulation was developed for magnetic nanoparticles in a liquid dielectric to investigate the AC breakdown voltage of the magnetic nanofluids according to the volume concentration of the magnetic nanoparticles. In prior research, we found that the dielectric breakdown voltage of the transformer oil-based magnetic nanofluids was positively or negatively affected according to the amount of magnetic nanoparticles under a testing condition of dielectric fluids, and the trajectory of the magnetic nanoparticles in a fabricated chip was visualized to verify the related phenomena via measurements and computations. In this study, a numerical simulation of magnetic nanoparticles in an insulating fluid was developed to model particle tracing for AC breakdown mechanisms happened to a sphere-sphere electrode configuration and to propose a possible mechanism regarding the change in the breakdown strength due to the behavior of the magnetic nanoparticles with different applied voltages.

Monitoring 3D dose distributions in proton therapy by reconstruction using an iterative method.

The Bragg peak of protons can be determined by measuring prompt γ-rays. In this study, prompt γ-rays detected by single-photon emission computed tomography with a geometrically optimized collimation system were reconstructed by an iterative method. The falloff position by iterative method (52.48mm) was most similar to the Bragg peak (52mm) of an 80MeV proton compared with those of back-projection (54.11mm) and filtered back-projection (54.91mm) methods. Iterative method also showed better image performance than other methods.

RNA sequencing identifies novel markers of non-small cell lung cancer.

INTRODUCTION: The development of reliable gene expression profiling technology increasingly impacts our understanding of lung cancer biology. Here, we used RNA sequencing (RNA-Seq) to compare the transcriptomes of non-small cell lung cancer (NSCLC) and normal lung tissues and to investigate expression in lung cancer tissues. METHODS: We enrolled 88 male patients (mean age, 61.2 years) with NSCLC. RNA-Seq was performed on 88 pairs of NSCLC tumor tissue and non-tumor tissue from 54 patients with adenocarcinoma and 34 patients with squamous cell carcinoma. Immunohistochemistry was performed to validate differential candidate gene expression in a different NSCLC group. RESULTS: RNA-Seq produced 25.41 × 10(6) (± 8.90 × 10(6)) reads in NSCLC tissues and 24.70×10(6) (± 4.70 × 10(6)) reads in normal lung tissues [mean (± standard deviation)]. Among the genes expressed in both tissues, 335 were upregulated and 728 were downregulated ≥ 2-fold (p < 0.001). Four upregulated genes - CBX3, GJB2, CRABP2, and DSP - not previously reported in lung cancer were studied further. Their altered expression was verified by immunohistochemistry in a different set of NSCLC tissues (n = 154). CBX3 was positive in 90.3% (139 cases) of the samples; GJB2, in 22.7% (35 cases); CRABP2, in 72.1% (111 cases); and DSP, in 17.5% (27 cases). The positive rate of CRABP2 was higher in adenocarcinoma than squamous cell carcinoma (p < 0.01). CONCLUSIONS: CBX3 and CRABP2 expression was markedly increased in lung cancer tissues and especially CRABP2 may be promising candidate genes in lung adenocarcinoma.

Primary aortoenteric fistula to the sigmoid colon in association with intra-abdominal abscess.

Primary aortoenteric fistula (PAEF) is a rare but catastrophic cause of massive gastrointestinal bleeding. Diagnosis of PAEF is difficult to make and is frequently delayed without strong clinical suspicion. Timely surgical intervention is essential for patient's survival. We report on a case of an 86-year-old woman with no history of abdominal surgery, who presented with abdominal pain. Initially, computed tomography scan showed an intra-abdominal abscess, located anterior to the aortic bifurcation. However, she was discharged without treatment because of spontaneous improvement on a follow-up computed tomography scan, which showed a newly developed right common iliac artery aneurysm. One week later, she was readmitted due to recurrent abdominal pain. On the second day of admission, sudden onset of gastrointestinal bleeding occurred for the first time. After several endoscopic examinations, an aortoenteric fistula bleeding site was found in the sigmoid colon, and aortography showed progression of a right common iliac artery aneurysm. We finally concluded that intra-abdominal abscess induced an infected aortic aneurysm and enteric fistula to the sigmoid colon. This case demonstrated an extremely rare type of PAEF to the sigmoid colon caused by an infected abdominal aortic aneurysm, which has rarely been reported.

Nicotine dependence as a moderator of genetic influences on smoking cessation treatment outcome.

BACKGROUND: Genetic influences on smoking cessation treatment outcome may be affected by pretreatment patient characteristics. Nicotine dependence is arguably the most salient clinical factor in smoking cessation. METHODS: In this secondary analysis of clinical trial data (N=793), we examined nicotine dependence severity as a moderator of the effects of 1198 single nucleotide polymorphisms (SNPs) in 53 biologically-relevant gene regions on smoking cessation outcomes. P-values were adjusted to account for multiple correlated SNPs within a gene region; corrected system-wide significance was 5 × 10(-4). RESULTS: SNP × nicotine dependence interactions reached region-wide significance for several SNPs in the Dopamine Beta Hydroxylase (DBH) locus (0.0005

Nonalcoholic fatty liver disease is associated with coronary artery disease in Koreans.

AIM: To investigate whether nonalcoholic fatty liver disease (NAFLD) affects coronary artery disease (CAD) and identify candidate mediators. METHODS: Patients who underwent coronary angiography were consecutively recruited. The patients were classified into four groups by coronary artery stenosis: A, insignificant; B, one-vessel disease; C, two-vessel disease; and D, three-vessel disease. Abdominal ultrasonography was performed to determine the presence of a fatty liver and categorize by grade: 0, no evidence; 1, mild; 2, moderate; and 3, severe. We measured not only known CAD risk factors, but also serum insulin, HOMA-index, adiponectin, interleukin-6, tumor necrosis factor-α and high-sensitivity C-reactive protein levels. RESULTS: Of the 134 patients who met the inclusion criteria, 82 (61.2%) had ultrasonographically diagnosed NAFLD. Among the 46 patients with CAD, 37 (80.4%) had evidence of a fatty liver. The two groups (A vs B-D) were significantly different in terms of age, total cholesterol, triglycerides, low-density lipoprotein levels and fatty liver. Coronary artery stenosis was strongly associated with fatty liver in a grade-dependent manner (P = 0.025). In binary logistic regression, NAFLD was a significant independent predictor of CAD (P = 0.03, OR = 1.685; 95%CI: 1.051-2.702). Among the candidate mediators, the serum adiponectin level showed a trend toward lowering based on CAD progression (P = 0.071). CONCLUSION: NAFLD is an independent risk factor for CAD in a grade-dependent manner. Moreover, adiponectin might be related to the pathogenesis of NAFLD.

A trachea-inspired bifurcated microfilter capturing viable circulating tumor cells via altered biophysical properties as measured by atomic force microscopy.

Circulating tumor cells (CTCs), though exceedingly rare in the blood, are nonetheless becoming increasingly important in cancer diagnostics. Despite this keen interest and the growing number of potential clinical applications, there has been limited success in developing a CTC isolation platform that simultaneously optimizes recovery rates, purity, and cell compatibility. Herein, a novel tracheal carina-inspired bifurcated (TRAB) microfilter system is reported, which uses an optimal filter gap size satisfying both 100% theoretical recovery rate and purity, as determined by biomechanical analysis and fluid-structure interaction (FSI) simulations. Biomechanical properties are also used to clearly discriminate between cancer cells and leukocytes, whereby cancer cells are selectively bound to melamine microbeads, which increase the size and stiffness of these cells. Nanoindentation experiments are conducted to measure the stiffness of leukocytes as compared to the microbead-conjugated cancer cells, with these parameters then being used in FSI analyses to optimize the filter gap size. The simulation results show that given a flow rate of 100 µL min(-1), an 8 µm filter gap optimizes the recovery rate and purity. MCF-7 breast cancer cells with solid microbeads are spiked into 3 mL of whole blood and, by using this flow rate along with the optimized microfilter dimensions, the cell mixture passes through the TRAB filter, which achieves a recovery rate of 93% and purity of 59%. Regarding cell compatibility, it is verified that the isolation procedure does not adversely affect cell viability, thus also confirming that the re-collected cancer cells can be cultured for up to 8 days. This work demonstrates a CTC isolation technology platform that optimizes high recovery rates and cell purity while also providing a framework for functional cell studies, potentially enabling even more sensitive and specific cancer diagnostics.