Immune evasion in HPV− head and neck precancer–cancer transition is driven by an aneuploid switch involving chromosome 9p loss

An aneuploid-immune paradox encompasses somatic copy-number alterations (SCNAs), unleashing a cytotoxic response in experimental precancer systems, while conversely being associated with immune suppression and cytotoxic-cell depletion in human tumors, especially head and neck cancer (HNSC). We present evidence from patient samples and cell lines that alterations in chromosome dosage contribute to an immune hot-to-cold switch during human papillomavirus-negative (HPV⁻) head and neck tumorigenesis. Overall SCNA (aneuploidy) level was associated with increased CD3⁺ and CD8⁺ T cell microenvironments in precancer (mostly CD3⁺, linked to trisomy and aneuploidy), but with T cell-deficient tumors. Early lesions with 9p21.3 loss were associated with depletion of cytotoxic T cell infiltration in TP53 mutant tumors; and with aneuploidy were associated with increased NK-cell infiltration. The strongest driver of cytotoxic T cell and Immune Score depletion in oral cancer was 9p-arm level loss, promoting profound decreases of pivotal IFN-γ-related chemokines (e.g., CXCL9) and pathway genes. Chromosome 9p21.3 deletion contributed mainly to cell-intrinsic senescence suppression, but deletion of the entire arm was necessary to diminish levels of cytokine, JAK-STAT, and Hallmark NF-κB pathways. Finally, 9p arm-level loss and JAK2-PD-L1 codeletion (at 9p24) were predictive markers of poor survival in recurrent HPV⁻ HNSC after anti–PD-1 therapy; likely amplified by independent aneuploidy-induced immune-cold microenvironments observed here. We hypothesize that 9p21.3 arm-loss expansion and epistatic interactions allow oral precancer cells to acquire properties to overcome a proimmunogenic aneuploid checkpoint, transform and invade. These findings enable distinct HNSC interception and precision-therapeutic approaches, concepts that may apply to other CN-driven neoplastic, immune or aneuploid diseases, and immunotherapies.

The genetic bases for predisposition, and neoplastic transformation, to cancer have been increasingly well described. However, it remains less clear how early, precancer cells employ these genetic alterations to acquire the characteristic features and properties (1) of malignant disease. For example, studies of the immune landscape led to breakthrough trials of programmed death-1 (PD-1) inhibitors for recurrent, metastatic head and neck squamous cell carcinoma (HNSC) therapy (2–4). This underscores the importance of immune modulation in these tumors, despite a still suboptimal overall response rate of less than 20% in advanced cancers. Immune response within tumors has been observed to be strongest at the earliest neoplastic stages, as reported recently in lung adenocarcinoma precursors (5). As such, new, immune-based strategies could be developed to reduce the high global burden of HNSC, by intercepting the most common precursor of the most common HNSC presentation: HPV⁻ oral squamous cell carcinomas (6–8).Studies of chromosome somatic copy-number (CN) alteration (SCNA) profiles have reported the impact of 3p14, 9p21, or 17p13 loss in molecular models of HNSC progression (9) and risk (10–15). Early studies reported that patients with oral precancers harboring 9p21 and/or 3p14 loss were at significantly greater cancer risk than those with retention at these loci (10, 16). A comprehensive, prospective validation study examined the relative contribution of six candidate chromosome-arm regions. 9p21 loss had the greatest influence on cancer risk (13). The mechanism underlying the association between CN and malignant transformation of precancers, however, is unclear (17–20). Studies of CN-altered neoplastic cells have shown that SCNAs can trigger a cytotoxic response in experimental precancer systems (21, 22) but, paradoxically, were associated with immune evasion (23) and suppression (24) in computational studies of naturally occurring human cancers. The latter, in melanoma, found that nonresponders to PD-1 and CTLA-4 blockade had higher CN alteration and loss burdens, which correlated with immunologically cold tumors, characterized by cytotoxic-cell, marker, and metric reductions, and suppressive microenvironment cell, network, and signal increases (23–26). This SCNA-cold association was particularly strong in our previous, pan-The Cancer Genome Atlas (TCGA) computational study in HNSC (23). These data point to a putative in vivo switch from immune hot-to-cold in the precancer–cancer transition, and raise the hypothesis that SCNAs in precursor lesions contribute to malignant transformation through genomic events and mechanisms that enable the acquisition of immune-suppressive, evasive properties. To test this hypothesis, we evaluated CN influence on immune profiles and outcomes in a large prospective oral precancer patient cohort, and HPV⁻ HNSC (tissue specimens and cell lines) and anti–PD-1–treated recurrent-disease cohorts.     

Hidden Genetic Variation in LCA9‐Associated Congenital Blindness Explained by 5′UTR Mutations and Copy‐Number Variations of NMNAT1

Leber congenital amaurosis (LCA) is a severe autosomal‐recessive retinal dystrophy leading to congenital blindness. A recently identified LCA gene is NMNAT1, located in the LCA9 locus. Although most mutations in blindness genes are coding variations, there is accumulating evidence for hidden noncoding defects or structural variations (SVs). The starting point of this study was an LCA9‐associated consanguineous family in which no coding mutations were found in the LCA9 region. Exploring the untranslated regions of NMNAT1 revealed a novel homozygous 5′UTR variant, c.‐70A>T. Moreover, an adjacent 5′UTR variant, c.‐69C>T, was identified in a second consanguineous family displaying a similar phenotype. Both 5′UTR variants resulted in decreased NMNAT1 mRNA abundance in patients’ lymphocytes, and caused decreased luciferase activity in human retinal pigment epithelial RPE‐1 cells. Second, we unraveled pseudohomozygosity of a coding NMNAT1 mutation in two unrelated LCA patients by the identification of two distinct heterozygous partial NMNAT1 deletions. Molecular characterization of the breakpoint junctions revealed a complex Alu‐rich genomic architecture. Our study uncovered hidden genetic variation in NMNAT1‐associated LCA and emphasized a shift from coding to noncoding regulatory mutations and repeat‐mediated SVs in the molecular pathogenesis of heterogeneous recessive disorders such as hereditary blindness.     

A virtual pointer to support the adoption of professional vision in laparoscopic training

Purpose

To assess a virtual pointer in supporting surgical trainees’ development of professional vision in laparoscopic surgery.

Methods

We developed a virtual pointing and telestration system utilizing the Microsoft Kinect movement sensor as an overlay for any imagine system. Training with the application was compared to a standard condition, i.e., verbal instruction with un-mediated gestures, in a laparoscopic training environment. Seven trainees performed four simulated laparoscopic tasks guided by an experienced surgeon as the trainer. Trainee performance was subjectively assessed by the trainee and trainer, and objectively measured by number of errors, time to task completion, and economy of movement.

Results

No significant differences in errors and time to task completion were obtained between virtual pointer and standard conditions. Economy of movement in the non-dominant hand was significantly improved when using virtual pointer (\(p = 0.012\)). The trainers perceived a significant improvement in trainee performance in virtual pointer condition (\(p < 0.001\)), while the trainees perceived no difference. The trainers’ perception of economy of movement was similar between the two conditions in the initial three runs and became significantly improved in virtual pointer condition in the fourth run (\(p = 0.017\)).

Conclusions

Results show that the virtual pointer system improves the trainer’s perception of trainee’s performance and this is reflected in the objective performance measures in the third and fourth training runs. The benefit of a virtual pointing and telestration system may be perceived by the trainers early on in training, but this is not evident in objective trainee performance until further mastery has been attained. In addition, the performance improvement of economy of motion specifically shows that the virtual pointer improves the adoption of professional vision— improved ability to see and use laparoscopic video results in more direct instrument movement.

     

Enhancing social networks: a qualitative study of health and social care practice in UK mental health services

People with severe mental health problems such as psychosis have access to less social capital, defined as resources within social networks, than members of the general population. However, a lack of theoretically and empirically informed models hampers the development of social interventions which seek to enhance an individual's social networks. This paper reports the findings of a qualitative study, which used ethnographic field methods in six sites in England to investigate how workers helped people recovering from psychosis to enhance their social networks. This study drew upon practice wisdom and lived experience to provide data for intervention modelling. Data were collected from 73 practitioners and 51 people who used their services in two phases. Data were selected and coded using a grounded theory approach to depict the key themes that appeared to underpin the generation of social capital within networks. Findings are presented in four over‐arching themes – worker skills, attitudes and roles; connecting people processes; role of the agency; and barriers to network development. The sub‐themes which were identified included worker attitudes; person‐centred approach; equality of worker–individual relationship; goal setting; creating new networks and relationships; engagement through activities; practical support; existing relationships; the individual taking responsibility; identifying and overcoming barriers; and moving on. Themes were consistent with recovery models used within mental health services and will provide the basis for the development of an intervention model to enhance individuals’ access to social capital within networks.     

HIV Infection Among People Who Inject Drugs in the United States: Geographically Explained Variance Across Racial and Ethnic Groups

Objectives. We explored how variance in HIV infection is distributed across multiple geographical scales among people who inject drugs (PWID) in the United States, overall and within racial/ethnic groups.Methods. People who inject drugs (n = 9077) were recruited via respondent-driven sampling from 19 metropolitan statistical areas (MSAs) for the Centers for Disease Control and Prevention’s 2009 National HIV Behavioral Surveillance system. We used multilevel modeling to determine the percentage of variance in HIV infection explained by zip codes, counties, and MSAs where PWID lived, overall and for specific racial/ethnic groups.Results. Collectively, zip codes, counties, and MSAs explained 29% of variance in HIV infection. Within specific racial/ethnic groups, all 3 scales explained variance in HIV infection among non-Hispanic/Latino White PWID (4.3%, 0.2%, and 7.5%, respectively), MSAs explained variance among Hispanic/Latino PWID (10.1%), and counties explained variance among non-Hispanic/Latino Black PWID (6.9%).Conclusions. Exposure to potential determinants of HIV infection at zip codes, counties, and MSAs may vary for different racial/ethnic groups of PWID, and may reveal opportunities to identify and ameliorate intraracial inequities in exposure to determinants of HIV infection at these geographical scales.Since the mid-1990s, there has been an increase in studies evaluating whether features of the social, economic, physical, and political environment (i.e., place characteristics) affect health. This focus on place characteristics is evident in the development of theories conceptualizing place characteristics as health determinants,1–3 in the use of geospatial and systematic social observation methods to measure place characteristics,4–10 in the application of multilevel modeling to assess the potential impacts of place characteristics,11–18 and in the recognition that interventions should not solely encourage individual behavior change but also modify environmental features.3,16,19Literature emerging from this field of research demonstrates that place characteristics operationalized at different geographical scales influence psychosocial processes and individual behaviors that increase vulnerability to several health outcomes. With rare exception,20–24 however, studies of place and health typically assess the potential influence of place characteristics at a single geographical scale and do not simultaneously evaluate characteristics of other geographical scales. For example, several studies, including our own,25,26 sample participants from a single metropolitan statistical area (MSA) to assess the relationships of census tract characteristics to health, without sampling participants from multiple MSAs to simultaneously assess the relationships of tract-, county-, and MSA-level characteristics to health.25–32 The decision to focus on characteristics of a single geographical scale may arise because of data availability, cost constraints, or feasibility.Studies of place and health that focus on a single geographical scale, however, may misspecify relationships and hinder the exploration of causal pathways in 2 ways. First, studies that focus on features measured at a single geographical scale may overlook potential health determinants that are operationalized at other geographical scales. For instance, research assessing the relationships of features of neighborhoods (e.g., economic deprivation, racial/ethnic composition, policing practices, and “crackdowns”) cannot determine the influence of policies, laws, and governmental expenditures that are operationalized at county, MSA, and state levels, and shape neighborhood environments. Second, studies of features of a single geographical scale cannot determine whether relationships between characteristics operating at one geographical scale are confounded, mediated, or modified by characteristics of other geographic scales.3,16,33 The possibility that at least 1 of these mechanisms can occur has been demonstrated in research conducted by Warner and Gomez, which suggests that, among Black women diagnosed with breast cancer, residing in census blocks with high concentrations of Black residents is more protective against mortality in more racially segregated metropolitan areas than less racially segregated metropolitan areas.34In addition, research assessing the association of place-based factors with health outcomes rarely highlights the extent to which variance in health outcomes is explained by place and place-based factors. Determining whether health outcomes vary geographically can generate hypotheses about inequities in exposure to potential place-based determinants of health, and thereby inform how interventions and social policies are developed and spatially concentrated.35The present study illustrates the generative possibilities of extending research beyond a single geographical scale by achieving 2 primary aims. The study’s first aim is to determine the share of total variance in HIV infection that is apportioned to zip codes, counties, and MSAs among people who inject drugs (PWID). In the United States, PWID account for 22% of people living with HIV,36 and a growing body of literature demonstrates that features of neighborhoods such as census-tract racial composition and block-level social or physical disorder are associated with HIV-related outcomes among PWID,37,38 as are features of MSAs, including drug-related law enforcement, income inequality, residential segregation, and health service access.39–41 Revealing the geographical scale to which variance in HIV infection is apportioned among PWID can stimulate hypotheses about inequities in exposure to place-based determinants of HIV and inform the development and tailoring of place-based interventions. For example, finding high MSA-level variance in HIV infection may support analyses of whether MSA-level variations in health care service access predict variance in HIV serostatus and, if they do, support interventions to increase health care access in low-access MSAs. In contrast, if little to no variance in HIV infection among PWID is apportioned to MSAs, PWID may encounter a relatively uniform exposure to health care service access.Previous studies have found that variance in some health outcomes vary across racial/ethnic groups.42,43 The second aim of this study therefore tests the hypothesis that variance in HIV infection will differ within each of 3 racial/ethnic groups of PWID: non-Hispanic/Latino Whites, non-Hispanic/Latino Blacks, and Hispanics/Latinos.     

Utility Values Associated with Diabetic Retinopathy in Chennai,India

Abstract

Purpose: The aim of this study was to estimate utility values associated with different severity stages of diabetic retinopathy (DR) in India by a direct elicitation method (time-trade off, TTO) and indirectly by questionnaire.

Methods: People with diabetes aged 40 years and over were recruited from an on-going DR epidemiology study and a laser clinic in Chennai, India. Utility values were elicited using the direct TTO method and indirectly through a validated questionnaire (EQ-5D).

Results: Of 249 participants, 30 had no DR, 73 had non-proliferative DR, 114 had sight-threatening DR, and 32 were blind from DR (bilateral visual acuity <6/60). The mean TTO utility value was 0.73 (standard deviation, SD, 0.31). TTO utility values decreased with increasing severity of DR (p?<?0.001) and were significantly lower among participants with sight threatening DR (0.70, SD 0.33) and blindness (0.55, SD 0.24) compared to those with no DR (0.89, SD 0.25) after adjustment for sociodemographic and clinical factors. Blindness from DR was independently associated with a lower EQ-5D utility value. The utility value derived from EQ-5D (0.06) associated with being blind from DR was substantially lower than that of the TTO utility value (0.55).

Conclusions: This study provides estimates of utility values that can be used in economic evaluations of DR screening strategies in India. The relatively low utility values associated with blindness highlights the importance of screening programs for early detection of the sight-threatening stages to prevent vision loss from DR in this setting.     

Punch's use of Graphic Satire for Middle Class Scientific Engagement, 1889–1914

Punch was the foremost satirical publication of the nineteenth and early twentieth century, engaging thoughtfully and seriously with the world of science. This article examines three of Punch's graphic satires concerning medical innovation. As Punch relied heavily on topical humour, many of its satires can trace their roots to occurrences reported in the popular press, which help to put the cartoons in context for an accurate reading of Punch's motives.

Punch's method, to educate and engage its loyal readers in debates about the role of medicine and doctors whilst making them laugh, would seem to be unique.