Evaluation of variable new antigen receptors (vNARs) as a novel cathepsin S (CTSS) targeting strategy.

Aberrant activity of the cysteine protease Cathepsin S (CTSS) has been implicated across a wide range of pathologies. Notably in cancer, CTSS has been shown to promote tumour progression, primarily through facilitating invasion and migration of tumour cells and augmenting angiogenesis. Whilst an attractive therapeutic target, more efficacious CTSS inhibitors are required. Here, we investigated the potential application of Variable New Antigen Receptors (vNARs) as a novel inhibitory strategy. A panel of potential vNAR binders were identified following a phage display panning process against human recombinant proCTSS. These were subsequently expressed, purified and binding affinity confirmed by ELISA and SPR based approaches. Selected lead clones were taken forward and were shown to inhibit CTSS activity in recombinant enzyme activity assays. Further assessment demonstrated that our lead clones functioned by a novel inhibitory mechanism, by preventing the activation of proCTSS to the mature enzyme. Moreover, using an intrabody approach, we exhibited the ability to express these clones intracellularly and inhibit CTSS activity whilst lead clones were also noted to impede cell invasion in a tumour cell invasion assay. Collectively, these findings illustrate a novel mechanistic approach for inhibiting CTSS activity, with anti-CTSS vNAR clones possessing therapeutic potential in combating deleterious CTSS activity. Furthermore, this study exemplifies the potential of vNARs in targeting intracellular proteins, opening a range of previously "undruggable" targets for biologic-based therapy.

Single-cell mapping identifies MSI+ cells as a common origin for diverse subtypes of pancreatic cancer.

Identifying the cells from which cancers arise is critical for understanding the molecular underpinnings of tumor evolution. To determine whether stem/progenitor cells can serve as cells of origin, we created a Msi2-CreERT2 knock-in mouse. When crossed to CAG-LSL-MycT58A mice, Msi2-CreERT2 mice developed multiple pancreatic cancer subtypes: ductal, acinar, adenosquamous, and rare anaplastic tumors. Combining single-cell genomics with computational analysis of developmental states and lineage trajectories, we demonstrate that MYC preferentially triggers transformation of the most immature MSI2+ pancreas cells into multi-lineage pre-cancer cells. These pre-cancer cells subsequently diverge to establish pancreatic cancer subtypes by activating distinct transcriptional programs and large-scale genomic changes, and enforced expression of specific signals like Ras can redirect subtype specification. This study shows that multiple pancreatic cancer subtypes can arise from a common pool of MSI2+ cells and provides a powerful model to understand and control the programs that shape divergent fates in pancreatic cancer.

Smarcd3 is an epigenetic modulator of the metabolic landscape in pancreatic ductal adenocarcinoma.

Pancreatic cancer is characterized by extensive resistance to conventional therapies, making clinical management a challenge. Here we map the epigenetic dependencies of cancer stem cells, cells that preferentially evade therapy and drive progression, and identify SWI/SNF complex member SMARCD3 as a regulator of pancreatic cancer cells. Although SWI/SNF subunits often act as tumor suppressors, we show that SMARCD3 is amplified in cancer, enriched in pancreatic cancer stem cells and upregulated in the human disease. Diverse genetic mouse models of pancreatic cancer and stage-specific Smarcd3 deletion reveal that Smarcd3 loss preferentially impacts established tumors, improving survival especially in context of chemotherapy. Mechanistically, SMARCD3 acts with FOXA1 to control lipid and fatty acid metabolism, programs associated with therapy resistance and poor prognosis in cancer. These data identify SMARCD3 as an epigenetic modulator responsible for establishing the metabolic landscape in aggressive pancreatic cancer cells and a potential target for new therapies.

Intra-dialytic intracranial pressure monitoring in a patient with lumbo-peritoneal shunt for idiopathic intracranial hypertension.

We present the case of a 25-year-old female with End-Stage Renal Disease (ESRD) and Idiopathic Intracranial Hypertension (IIH) who developed severe headaches during haemodialysis (HD). The headaches resolved several hours after each HD session. We were able to diagnose dialysis disequilibrium syndrome (DDS) following intracranial pressure (ICP) monitoring and use a novel strategy to treat her symptoms.

From heavy cannabis use to psychosis: is it time to take action?

Cannabis is one of the most widely used recreational drugs among people with clinical psychosis, after nicotine and alcohol. There has been a debate in psychiatry about whether or not we can infer a cause-and-effect relationship between the use of cannabis and psychotic disorders. In this editorial, we first present and critically discuss the evidence to date of the association between heavy cannabis use and psychosis. We argue that while the biological mechanisms underlying individual susceptibility to develop a psychotic disorder following heavy cannabis use are still unknown, heavy cannabis use remains the most modifiable risk factor for the onset of psychotic disorders and for its clinical and functional outcome. This demands a clear move towards both primary and secondary prevention intervention to reduce the impact of heavy cannabis use on the incidence and prevalence of psychotic disorders.

The Role of the Microenvironment and Immune System in Regulating Stem Cell Fate in Cancer.

Despite gains in knowledge of the intrinsic signals governing cancer progression, effective clinical management of cancer remains a challenge. Drug resistance and relapse, pose the greatest barriers to cancer care, and are often driven by the co-option of stem cell programs by subpopulations of aggressive cancer cells. Here, we focus on the role of the microenvironment in the acquisition and/or maintenance of stem cell states in cancer in the context of resistance and metastasis. We further discuss the role of cancer stem cells in immune evasion through the course of metastasis, dormancy, and relapse. Understanding the niche in which cancer stem cells live and the signals that sustain them may lead to new strategies that target them by disrupting microenvironmental support.

T-cell pseudolymphoma secondary to ixazomib for multiple myeloma.

We present a case of a 54-year-old male with multiple myeloma (MM) who presented with widespread pruritic erythematous lesions following ixazomib treatment. This occurred after his third cycle of treatment with ixazomib, thalidomide and dexamethasone and was controlled by potent steroids and temporary cessation of ixazomib. The strong correlation between the timeline of the rash, ixazomib treatment and subsequent cessation led to a diagnosis of a drug-induced rash. Skin biopsy histology, immunochemistry and the absence of monoclonal T-cell receptor gene rearrangement further confirmed the diagnosis of a T-cell pseudolymphoma secondary to ixazomib. Ixazomib is an oral proteasome inhibitor used in the treatment of MM. Other proteasome inhibitors have been reported to trigger cutaneous adverse effects. However, to our knowledge, this is the first report of pseudolymphoma following proteasome inhibitor use. Dermatologists should be aware of this potential effect and the possible management pathways such as cessation and dose reduction.

Extensive mucocutaneous, oesophageal and otic lichen planus secondary to nivolumab therapy.

We report a 73-year-old female with metastatic renal cell carcinoma who developed a widespread lichenoid reaction following nivolumab treatment. The timeline of the reaction strongly correlated with the nivolumab treatment and subsequent cessation. Our patient had cutaneous, mucosal, otic, ophthalmic and oesophageal involvement, demonstrating the potentially extensive nature of lichenoid reactions to anti-programmed cell death receptor-1 (anti-PD1) therapies. Although lichenoid reactions to anti-PD1 therapies are now well recognized, there have been no previous reports of otic or oesophageal involvement in the literature. Although cutaneous lichenoid reactions do not tend to be severe or treatment limiting, more widespread systemic lichenoid reactions are challenging to manage, particularly in the context of malignancy. This very unusual case highlights the importance of considering involvement beyond the skin in all lichenoid skin reactions.

Skin cancer multiplicity in lung transplant recipients: a prospective population-based study.

BACKGROUND: Lung transplant recipients are at high risk of skin cancer, but precise annual incidence rates of treated skin cancers per patient are unknown. OBJECTIVES: To perform a prospective assessment of the total burden of histologically confirmed squamous cell carcinoma (SCC) and basal cell carcinoma (BCC) and associated factors in lung transplant recipients. METHODS: A population-based cohort of 125 Queensland lung transplant recipients aged 18 years and over, recruited between 2013 and 2015, were followed to the end of 2016. All underwent dermatological skin examinations at baseline and annually thereafter and patients self-reported all interim treated skin cancers, which were verified against pathology databases. Standard skin cancer risk factors were obtained via questionnaire, and details of medications were acquired from hospital records. RESULTS: During a median follow-up time of 1·7 years, 29 (23%) and 30 (24%) lung transplant recipients with a median duration of immunosuppression of 3·3 years developed SCC and BCC, respectively. The general population age-standardized incidence rates of SCC and BCC were 201 and 171 per 1000 person-years, respectively (based on first primary SCC or BCC during follow-up); however, on accounting for multiple primary tumours, corresponding incidence rates were 447 and 281 per 1000 person-years. Risk of multiple SCCs increased around sixfold in those aged ≥ 60 years and in those with previous skin cancer, and increased around threefold in those treated with the antifungal medication voriconazole. Multiple BCC risk rose threefold from age 60 years and tenfold for patients with previous skin cancer. CONCLUSIONS: Lung transplant recipients have very high incidence of multiple primary skin cancers. Close surveillance and assiduous prevention measures are essential. Linked Comment: Proby and Harwood. Br J Dermatol 2020; 183:416-417.

Untargeted mass spectrometry-based metabolomics approach unveils molecular changes in raw and processed foods and beverages.

In our daily lives, we consume foods that have been transported, stored, prepared, cooked, or otherwise processed by ourselves or others. Food storage and preparation have drastic effects on the chemical composition of foods. Untargeted mass spectrometry analysis of food samples has the potential to increase our chemical understanding of these processes by detecting a broad spectrum of chemicals. We performed a time-based analysis of the chemical changes in foods during common preparations, such as fermentation, brewing, and ripening, using untargeted mass spectrometry and molecular networking. The data analysis workflow presented implements an approach to study changes in food chemistry that can reveal global alterations in chemical profiles, identify changes in abundance, as well as identify specific chemicals and their transformation products. The data generated in this study are publicly available, enabling the replication and re-analysis of these data in isolation, and serve as a baseline dataset for future investigations.

Evaluation of gastric motility in nine dogs before and after prophylactic laparoscopic gastropexy: a pilot study.

BACKGROUND: The purpose of this study was to evaluate the effect of a prophylactic laparoscopic gastropexy on gastric motility in healthy large-breed dogs. METHODS: This was a prospective pilot study with nine healthy client-owned dogs. Each dog was its own control. Gastric motility was evaluated before and after laparoscopic gastropexy. Dogs were fed a standard diet three weeks before and after surgery. Gastric motility was measured before and 3 weeks after surgery. A wireless motility capsule (WMC) was used to measure gastric pH, intragastric pressure, temperature, frequency of contractions, motility index (MI) and transit time. Non-parametric statistical analysis was used to compare the paired data. Clients were contacted for follow-up information 2 years postoperatively. RESULTS: Median frequency of gastric contractions was 1.3 (range, 0.6-1.9 contractions/min) before gastropexy and 1.0 (range, 0.3-2.6 contractions/min) after gastropexy (P = 0.820). Median MI was 49.2 (range, 23.7-96.6) before gastropexy and 28.1 (range, 12.2-148.9) after gastropexy (P = 0.652). Median gastric emptying time was 1140 (range, 486-1230 min) before gastropexy and 1110 (range, 306-2610 min) after gastropexy (P = 0.570). During the hour before the WMC passed through the pylorus, median MI was 72.2 (range, 48.2-549.3) before gastropexy and 52.9 (range, 15.20-322.8) after gastropexy (P = 0.734), and frequency of contractions was 1.1 (range, 0.9-4.1 contractions/min) before gastropexy and 1.2 (range, 0.5-3.0 contractions/min) after gastropexy (P = 0.652). CONCLUSION: Motility in the stomach did not change in healthy dogs after prophylactic laparoscopic gastropexy. We conclude that preventive laparoscopic gastropexy does not induce gastroparesis.

A Multiscale Map of the Stem Cell State in Pancreatic Adenocarcinoma.

Drug resistance and relapse remain key challenges in pancreatic cancer. Here, we have used RNA sequencing (RNA-seq), chromatin immunoprecipitation (ChIP)-seq, and genome-wide CRISPR analysis to map the molecular dependencies of pancreatic cancer stem cells, highly therapy-resistant cells that preferentially drive tumorigenesis and progression. This integrated genomic approach revealed an unexpected utilization of immuno-regulatory signals by pancreatic cancer epithelial cells. In particular, the nuclear hormone receptor retinoic-acid-receptor-related orphan receptor gamma (RORγ), known to drive inflammation and T cell differentiation, was upregulated during pancreatic cancer progression, and its genetic or pharmacologic inhibition led to a striking defect in pancreatic cancer growth and a marked improvement in survival. Further, a large-scale retrospective analysis in patients revealed that RORγ expression may predict pancreatic cancer aggressiveness, as it positively correlated with advanced disease and metastasis. Collectively, these data identify an orthogonal co-option of immuno-regulatory signals by pancreatic cancer stem cells, suggesting that autoimmune drugs should be evaluated as novel treatment strategies for pancreatic cancer patients.

Mechanisms underlying vulnerabilities after repeat mild traumatic brain injuries.

Traumatic brain injury (TBI) has drawn national attention for its high incidence and mechanistic complexity. The majority of TBI cases are "mild" in nature including concussions and mild TBI (mTBI). Concussions are a distinct form of mTBI where diagnosis is difficult, quantification of the incidence is challenging and there is greater risk for subsequent injuries. While concussions occur in the general population, it has become a hallmark injury consistently observed among adolescent and young adult athletes and the risks for repeat TBI (rTBI) is significant. Clinical and experimental evidence shows that the magnitude and duration of deficits is dependent on the number and the interval between injuries. Several studies suggest that metabolic vulnerabilities after injury may contribute to the window for cerebral vulnerability from rTBI. In addition to metabolism, this review addresses how age, sex and hormones also play an important role in the response to repeat concussions. Understanding how these factors collectively contribute to concussion and rTBI recovery is critically important in establishing age/sex appropriate return to play guidelines, injury prevention, therapeutic interventions and mitigation of long-term consequences of rTBI.

Promotion of the mediterranean diet incancer long-survivors by means of the Med-Food Anticancer Program: a pilot study.

BACKGROUND: The intervention "Med-Anticancer Food Program" has proven to be effective in promoting the Mediterranean Diet, significantly increasing the Mediterranean Adequacy Index in healthy subjects. There are no studies that have investigated the effectiveness of this intervention in individuals who have had a diagnosis of cancer. OBJECTIVE: To perform a pilot study to assess the opportunity of employing the methodology of the Med-Anticancer Food Program in order to encourage "long-term cancer survivors" to adhere to the Mediterranean Diet, as well as healthy people, and this in order to apply the program to larger groups. METHODS: From the residents' register of Foggia, a city in southern Italy, forty adults of both sexes, over 25 years of age, were recruited at random and assigned (1:1) as follows: - Twenty healthy subjects to the intervention-1 group - Twenty long-term cancer survivors to the intervention-2 group. The Med-Anticancer Food Program was applied to both groups with an articulated intervention 11 weeks long, followed by a 52-week period of follow up. By means of a food diary of the last 3 days, the Mediterranean Adequacy Index values were calculated before intervention (T0), after a period of 11 weeks of interventions (T1) and at the end of the 52 weeks of follow-up period (T2). The H0 hypothesis of the study was that there are no differences between the two interventions in reaching by T1, and maintaining at T2, values of Mediterranean Adequacy Index around 7, considered the optimum for adherence to the Mediterranean diet. RESULTS: Out of the subjects assigned to the intervention-1 group (n = 20), 11 subjects have completed the 52-months follow-up (55.0% ); for intervention-2, 16 (80%) out of 20 have completed it. The average age of subjects was 52.1 years. The Mediterranean Adequacy Index, of intervention-1 group significantly increased from 2.8 (T0) to 9.2 (T1) and to 9.0 (T2) (p <0.0001); whereas, in the intervention-2 group, Mediterranean Adequacy Index moved from 2.4 (T0) to 10.2 (T1) and to 9.3 (T2) (p <0.0001). The difference of Mediterranean Adequacy Index between the two study groups at T1 and T2 was not significant. Such non-significance persists also after the stratification by sex and age obtained with Mantel-Haenszel procedure. The performance of the values of the laboratory parameters considered (folic acid, total cholesterol, alkyl resorcinol) was similar in the subjects of both intervention 1 and 2, without any difference, while considered at a basal level T0, at T1 and at the end of the follow-up period (T2). CONCLUSIONS: The results of our work suggest the feasibility of conducting the Med-Anticancer Food Program in long-term cancer survivors. The results of the pilot study show that such intervention, carried on a small number of long term cancer survivors, is adequate to assess its feasibility but, due to the limited size of our study, a confirmation is required through larger nutritional prevention intervention studies.

Men across a range of ethnicities have a higher prevalence of diabetes: findings from a cross-sectional study of 500 000 UK Biobank participants.

AIMS: Studies show that white men have a higher prevalence of Type 2 diabetes mellitus than women at a given age and BMI, but equivalent standardized data for other ethnic groups in the UK are sparse. METHODS: This cross-sectional study analysed UK Biobank data from 489 079 participants to compare the prevalence of diabetes mellitus across four major ethnic groups including: 471 700 (96.4%) white, 7871 (1.6%) South Asian, 7974 (1.6%) black and 1534 (0.3%) Chinese participants, before and after standardizing for age, socio-economic status (SES), BMI and lifestyle factors including physical activity, TV viewing, fruit and vegetable intake, processed meat, red meat, oily fish, alcohol intake and smoking. A subgroup analysis of South Asians was also undertaken. RESULTS: Crude diabetes prevalence was higher in men across all four ethnicities. After standardizing for age, SES, BMI and lifestyle factors, a significant sex difference in diabetes prevalence persisted in white (men 6.0% vs. women 3.6%), South Asian (21.0% vs. 13.8%) and black individuals (13.3% vs. 9.7%) (P < 0.0001); there was a non-significant difference between Chinese men and women (7.1% vs. 5.5%) (P = 0.211). Sex differences persisted across South Asian subgroups. CONCLUSIONS: Men across a range of major ethnic groups including white, South Asian and black, have a higher prevalence of diabetes compared with women of similar age, BMI, SES and lifestyle in the UK.