Targeting a cytokine checkpoint enhances the fitness of armored cord blood CAR-NK cells.

Immune checkpoint therapy has resulted in remarkable improvements in the outcome for certain cancers. To broaden the clinical impact of checkpoint targeting, we devised a strategy that couples targeting of the cytokine-inducible Src homology 2-containing (CIS) protein, a key negative regulator of interleukin 15 (IL-15) signaling, with fourth-generation "armored" chimeric antigen receptor (CAR) engineering of cord blood-derived natural killer (NK) cells. This combined strategy boosted NK cell effector function through enhancing the Akt/mTORC1 axis and c-MYC signaling, resulting in increased aerobic glycolysis. When tested in a lymphoma mouse model, this combined approach improved NK cell antitumor activity more than either alteration alone, eradicating lymphoma xenografts without signs of any measurable toxicity. We conclude that targeting a cytokine checkpoint further enhances the antitumor activity of IL-15-secreting armored CAR-NK cells by promoting their metabolic fitness and antitumor activity. This combined approach represents a promising milestone in the development of the next generation of NK cells for cancer immunotherapy.

Evaluation of electronic pheromone trap capture conditions for Ips sexdentatus with climatic and temporal factors.

Controlling forest pests to maintain the sustainability of forests and ecosystem balance is one of the interests of modern forestry. In the evaluation of damage risks associated with forest pests, pheromone traps attract attention by providing early warnings. With the development of these traps in line with modern technology, more reliable data are obtained; these data are important in the identification and planning of pest management. In this study, a pheromone trap with electronic control unit was tested under field conditions. The capture of adult Ips sexdentatus under natural conditions during 103 days of the flying period was evaluated; 97.2% of the beetles captured in the trap were the target species. The comparison of the number of beetles recorded by the trap and manual counts revealed that the trap worked with an error margin of approximately 4%. However, no statistically significant difference was noted between these two counting methods. During the study, 59% of the total beetles were captured between May 27 and June 25. The average temperature at the period of the capture was 20.09 °C, average humidity was 66%, and average wind speed was 2.9 m/s. Of the captures, 73.9% occurred in the temperature range of 15-24.9 °C, 61.1% occurred in humidity range of 61-90%, 89.6% occurred at a wind speed of 0.3-5.4 m/s, and 77.3% occurred within the period from sunrise to sunset. When these four parameters were evaluated together, the most strongly associated parameter was daylight, followed by temperature, wind speed, and humidity.

A new design of electronic control unit involving microcontroller to determine important parameters for target species in forest.

Forests, a valuable source provided by nature to living beings, are indispensable for many living organisms; hence, it is important to ensure the sustainability of forests. Determining the factors that exposure threats to the forests, executing protective methods against them and putting these methods into practise are important for the ecological cycle. Bark beetles, which have destructive effects on the ecosystem, are one of the factors that expose a threat to forests. Therefore, monitoring of these species and determination of effective control strategies are increasingly gaining importance in forestry. Conventional pheromone traps, which are being currently used, provide limited information on flight times of target species. Therefore, the technological development of the capture systems of these traps will determine future control trends. Hence, pheromone traps with electronic control unit were prepared in earlier (ѵ1) and new designed (ѵ2) versions. In ѵ2, 97.5% of target species were counted, and instant temperature, humidity and time parameters at the time of capture were recorded at a practiced field work for the system. In addition to the instant parameters recorded in ѵ2, an anemometer used for measuring wind speed, which is considered to have influence on the behaviour of target species, was incorporated into the system. In the trials, the counting success rates under daylight and darkness conditions for Ips sexdentatus adults were 98.1 and 97%, whereas the counting success rates for Pityocteines curvidens adults, which are smaller in size, were 96 and 99%, respectively. In conclusion, data obtained by recording the amount of target species along with the capture moment and parameters related to this will be very useful and provide determinative in the management of target species.

A new approach to determine the capture conditions of bark beetles in pheromone-baited traps.

Forests form an organic unity with a great number of organic and inorganic components and tend to maintain the sustainability of their existing balance. However, some factors which adversely affect the balance of nature may interrupt this sustainability. The epidemic which is formed by bark beetles in their spreading region, due to various factors, changes the stability so much that interference is required. One of the most common methods used to monitor these beetles is pheromone-baited traps. The recognition of parameters, such as date (day/month/year), temperature and humidity, when bark beetles are captured in pheromone-baited traps, especially those used for monitoring will help to increase the trap efficiency on land and to develop an effective strategy for combating pests. In this study, an electronic control unit was added to pheromone-baited traps in order to obtain all of the above mentioned parameters. This unit operates with microcontrollers and data related to the parameters is saved in a storage unit. This is triggered by the beetle at the moment it is captured in the trap. A photovoltaic system was used to meet the energy needed for the system functioning and to complete the counting process in due time.

Leptomeningeal Metastasis from Non-Small Cell Lung Cancer and Current Landscape of Treatments.

Leptomeningeal metastasis (LM), also known as leptomeningeal carcinomatosis (LC), is a devastating complication of metastatic cancer that occurs when neoplastic cells invade the meningeal space. Diagnosis of LM remains challenging given the heterogeneous signs and symptoms at presentation and requires thorough neurological examination, cerebrospinal fluid (CSF) analysis, and MRI of the brain and spine with gadolinium. Detecting neoplastic cells in the CSF is the gold standard for diagnosing leptomeningeal metastases; however, it has low sensitivity and may require multiple CSF samples. New emerging technologies, such as liquid biopsy of CSF, have increased sensitivity and specificity for detecting circulating tumor cells in CSF. The management of LM in patients with NSCLC requires an individualized multidisciplinary approach. Treatment options include surgery for ventricular shunt placement, radiation therapy to bulky or symptomatic disease sites, systemic or intrathecal chemotherapy, molecularly targeted agents, and, more recently, immunotherapy. Targeting actionable mutations in LM from NSCLC, such as EGFR tyrosine kinase inhibitors or anaplastic lymphoma kinase gene rearrangement inhibitors, has shown encouraging results in terms of disease control and survival. Although there are limited data regarding the use of immunotherapy in LM, immunotherapy has produced promising results in several case reports. In this review, we focused on the epidemiology, pathophysiology, clinical presentation, diagnosis, and current treatment strategies, with a special emphasis on novel agents, including targeted therapies and immunotherapy of LM in patients with NSCLC.

Primary Hepatic Extra-gastrointestinal Stromal Tumors: Molecular Pathogenesis, Immunohistopathology, and Treatment.

Gastrointestinal stromal tumors are the most common mesenchymal tumors of the gastrointestinal tract. They originate from the interstitial cells of Cajal and are usually found in extrahepatic gastrointestinal sites. However, a small subset are derived from the liver and are known as primary hepatic gastrointestinal stromal tumors (PHGIST). They have a poor prognosis and are historically difficult to diagnose. Our objective was to review and update the latest evidence-based knowledge concerning PHGIST, with a focus on epidemiology, etiology, pathophysiology, clinical presentation, histopathology, and treatment. These tumors are usually found incidentally, occur sporadically, and are associated with mutations of KIT and PDGFRA genes. PHGIST is a diagnosis of exclusion, as it has the same molecular, immunochemistry and histological appearance as gastrointestinal stromal tumors (GIST). Thus, imaging, such as positron emission tomography-computed tomography (PET-CT) must be used to rule out metastatic GIST before a diagnosis can be made. However, with mutation analysis and pharmacological advances, tyrosine kinase inhibitors are typically pursued with or without surgical intervention. Other potential treatments include transcatheter arterial chemoembolization and tumor ablation. However, these are typically considered palliative options. As there are only a limited number of publications regarding PHGIST, data concerning morbidity and mortality are not yet available. Immunohistopathology can help develop screening guidelines and evaluating resistance to treatment.

Not a Statin-Induced Myopathy: Metastatic Pancreatic Adenocarcinoma Presenting As Paraneoplastic Myositis.

Polymyositis is an inflammatory disease that causes bilateral proximal muscle weakness; unlike dermatomyositis, it is not usually associated with malignancy. However, there are a handful of case reports documenting polymyositis in patients with lymphoma, breast, lung, and bladder cancer. Here we report a case of metastatic pancreatic adenocarcinoma disguised by presenting as polymyositis. Clinical presentation, laboratory values, muscle biopsy, and imaging were all diagnostic of paraneoplastic polymyositis. The patient has significantly improved in symptoms are receiving systemic steroids and pancreaticoduodenectomy.

Peritoneal Carcinomatosis as an Initial Presentation of Gastrointestinal Stromal Tumor: A Case Report and Review of the Literature.

Gastrointestinal stromal tumors (GISTs) are a rare type of tumor with a high risk of malignant transformation. The majority of GISTs are asymptomatic. Surgical resection remains the mainstay of treatment given that GIST is resistant to traditional chemotherapy and radiotherapy. In the last two decades, the discovery of targeted therapy with tyrosine kinase inhibitor therapy (TKI) and widespread mutation analysis of tumors have transformed the treatment of GIST. We present a case of a patient in whom imaging findings were consistent with carcinomatous peritonitis concerning a gynecological malignancy but who was later found to have an unresectable GIST which locally regressed with TKI.

Doege-Potter Syndrome with a Benign Solitary Fibrous Tumor: A Case Report and Literature Review.

Doege-Potter syndrome is a rare paraneoplastic syndrome that is often diagnosed incidentally during the workup of hypoglycemia of unclear etiology. It is characterized by a non-islet cell tumor hypoglycemia secondary to excessive production of partially processed IGF-II hormone from a solitary fibrous tumor (SFT). Often these tumors are intrathoracic, benign, and asymptomatic. Occasionally they present as a paraneoplastic event; hypertrophic osteoarthropathy in Pierre-Marie-Bamberger syndrome and hypoglycemia in Doege-Potter syndrome. The NAB2-STAT6 gene fusion is the hallmark of the SFT. Complete surgical resection of the tumor often results in resolution of symptoms and cure in most cases. Here we present the case of an 83-year-old non-diabetic female with recurrent syncopal events who was diagnosed with the Doege-Potter syndrome secondary to a SFT of pleura. Her tumor was positive for NAB2-STAT6 gene fusion on RT-PCR. Following the resection of the giant tumor mass, she became symptom-free within 24 h, and has remained asymptomatic at 4 months follow-up.

Targeting the αv integrin/TGF-ß axis improves natural killer cell function against glioblastoma stem cells.

Glioblastoma multiforme (GBM), the most aggressive brain cancer, recurs because glioblastoma stem cells (GSCs) are resistant to all standard therapies. We showed that GSCs, but not normal astrocytes, are sensitive to lysis by healthy allogeneic natural killer (NK) cells in vitro. Mass cytometry and single-cell RNA sequencing of primary tumor samples revealed that GBM tumor-infiltrating NK cells acquired an altered phenotype associated with impaired lytic function relative to matched peripheral blood NK cells from patients with GBM or healthy donors. We attributed this immune evasion tactic to direct cell-to-cell contact between GSCs and NK cells via αv integrin-mediated TGF-ß activation. Treatment of GSC-engrafted mice with allogeneic NK cells in combination with inhibitors of integrin or TGF-ß signaling or with TGFBR2 gene-edited allogeneic NK cells prevented GSC-induced NK cell dysfunction and tumor growth. These findings reveal an important mechanism of NK cell immune evasion by GSCs and suggest the αv integrin/TGF-ß axis as a potentially useful therapeutic target in GBM.

Large-scale GMP-compliant CRISPR-Cas9-mediated deletion of the glucocorticoid receptor in multivirus-specific T cells.

Virus-specific T cells have proven highly effective for the treatment of severe and drug-refractory infections after hematopoietic stem cell transplant (HSCT). However, the efficacy of these cells is hindered by the use of glucocorticoids, often given to patients for the management of complications such as graft-versus-host disease. To address this limitation, we have developed a novel strategy for the rapid generation of good manufacturing practice (GMP)-grade glucocorticoid-resistant multivirus-specific T cells (VSTs) using clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9) gene-editing technology. We have shown that deleting the nuclear receptor subfamily 3 group C member 1 (NR3C1; the gene encoding for the glucocorticoid receptor) renders VSTs resistant to the lymphocytotoxic effect of glucocorticoids. NR3C1-knockout (KO) VSTs kill their targets and proliferate successfully in the presence of high doses of dexamethasone both in vitro and in vivo. Moreover, we developed a protocol for the rapid generation of GMP-grade NR3C1 KO VSTs with high on-target activity and minimal off-target editing. These genetically engineered VSTs promise to be a novel approach for the treatment of patients with life-threatening viral infections post-HSCT on glucocorticoid therapy.

A novel immature natural killer cell subpopulation predicts relapse after cord blood transplantation.

Natural killer (NK) cells are highly heterogeneous, with vast phenotypic and functional diversity at the single-cell level. They are involved in the innate immune response against malignant and virus-infected cells. To understand the effect of NK diversity during immune recovery on the antitumor response after cord blood transplantation (CBT), we used high-dimensional mass cytometry and the metrics of NK cell diversity to study the NK cell repertoire in serial samples from 43 CBT recipients. A higher-diversity index based on single-cell combinatorial phenotypes was significantly associated with a lower risk for relapse after CBT (P = .005). Cytomegalovirus reactivation was a major factor in the development of a more diverse NK repertoire after CBT. Notably, we identified a group of patients whose CB-derived NK cells after transplantation possessed an immature phenotype (CB-NKim), characterized by poor effector function and a low diversity index. Frequencies of CB-NKim of 11.8% or higher during the early post-CBT recovery phase were highly predictive for relapse (area under the curve [AUC], 0.979), a finding that was validated in a second independent cohort of patients (n = 25; AUC, 0.977). Moreover, we showed that the maturation, diversity, and acquisition of effector function by CB-NKim early after CBT were driven by interleukin 15. Our data indicate that the diversity of the NK cell repertoire after CBT contributes importantly to the risk for subsequent relapse. We suggest that the use of diversity metrics and high-dimensional mass cytometry may be useful tools in predicting clinical outcomes and informing the design of therapeutic strategies to prevent relapse after CBT.