Identification, validation and biological characterisation of novel glioblastoma tumour microenvironment subtypes: implications for precision immunotherapy.

BACKGROUND: New precision medicine therapies are urgently required for glioblastoma (GBM). However, to date, efforts to subtype patients based on molecular profiles have failed to direct treatment strategies. We hypothesised that interrogation of the GBM tumour microenvironment (TME) and identification of novel TME-specific subtypes could inform new precision immunotherapy treatment strategies. MATERIALS AND METHODS: A refined and validated microenvironment cell population (MCP) counter method was applied to >800 GBM patient tumours (GBM-MCP-counter). Specifically, partition around medoids (PAM) clustering of GBM-MCP-counter scores in the GLIOTRAIN discovery cohort identified three novel patient clusters, uniquely characterised by TME composition, functional orientation markers and immune checkpoint proteins. Validation was carried out in three independent GBM-RNA-seq datasets. Neoantigen, mutational and gene ontology analysis identified mutations and uniquely altered pathways across subtypes. The longitudinal Glioma Longitudinal AnalySiS (GLASS) cohort and three immunotherapy clinical trial cohorts [treatment with neoadjuvant/adjuvant anti-programmed cell death protein 1 (PD-1) or PSVRIPO] were further interrogated to assess subtype alterations between primary and recurrent tumours and to assess the utility of TME classifiers as immunotherapy biomarkers. RESULTS: TMEHigh tumours (30%) displayed elevated lymphocyte, myeloid cell immune checkpoint, programmed cell death protein 1 (PD-1) and cytotoxic T-lymphocyte-associated protein 4 transcripts. TMEHigh/mesenchymal+ patients featured tertiary lymphoid structures. TMEMed (46%) tumours were enriched for endothelial cell gene expression profiles and displayed heterogeneous immune populations. TMELow (24%) tumours were manifest as an 'immune-desert' group. TME subtype transitions upon recurrence were identified in the longitudinal GLASS cohort. Assessment of GBM immunotherapy trial datasets revealed that TMEHigh patients receiving neoadjuvant anti-PD-1 had significantly increased overall survival (P = 0.04). Moreover, TMEHigh patients treated with adjuvant anti-PD-1 or oncolytic virus (PVSRIPO) showed a trend towards improved survival. CONCLUSIONS: We have established a novel TME-based classification system for application in intracranial malignancies. TME subtypes represent canonical 'termini a quo' (starting points) to support an improved precision immunotherapy treatment approach.

Impact of loss of NF-κB1, NF-κB2 or c-REL on SLE-like autoimmune disease and lymphadenopathy in Fas(lpr/lpr) mutant mice.

Defects in apoptosis can cause autoimmune disease. Loss-of-function mutations in the 'death receptor' FAS impair the deletion of autoreactive lymphocytes in the periphery, leading to progressive lymphadenopathy and systemic lupus erythematosus-like autoimmune disease in mice (Fas(lpr/lpr) (mice homozygous for the lymphoproliferation inducing spontaneous mutation)) and humans. The REL/nuclear factor-κB (NF-κB) transcription factors regulate a broad range of immune effector functions and are also implicated in various autoimmune diseases. We generated compound mutant mice to investigate the individual functions of the NF-κB family members NF-κB1, NF-κB2 and c-REL in the various autoimmune pathologies of Fas(lpr/lpr) mutant mice. We show that loss of each of these transcription factors resulted in amelioration of many classical features of autoimmune disease, including hypergammaglobulinaemia, anti-nuclear autoantibodies and autoantibodies against tissue-specific antigens. Remarkably, only c-REL deficiency substantially reduced immune complex-mediated glomerulonephritis and extended the lifespan of Fas(lpr/lpr) mice. Interestingly, compared with the Fas(lpr/lpr) animals, Fas(lpr/lpr)nfkb2(-/-) mice presented with a dramatic acceleration and augmentation of lymphadenopathy that was accompanied by severe lung pathology due to extensive lymphocytic infiltration. The Fas(lpr/lpr)nfkb1(-/-) mice exhibited the combined pathologies caused by defects in FAS-mediated apoptosis and premature ageing due to loss of NF-κB1. These findings demonstrate that different NF-κB family members exert distinct roles in the development of the diverse autoimmune and lymphoproliferative pathologies that arise in Fas(lpr/lpr) mice, and suggest that pharmacological targeting of c-REL should be considered as a strategy for therapeutic intervention in autoimmune diseases.

An exploration looking at the impact of domiciliary and day hospital delivery of stroke rehabilitation on informal carers.

OBJECTIVES: To explore the impact of two methods of post-hospital stroke rehabilitation on both carers' perceptions of the health services offered and their quality of life. SETTING: East Dorset Health Authority. SUBJECTS: Forty-six informal carers were recruited from a sample of 106, initially identified from stroke patients participating in a larger randomized controlled trial. DESIGN: Qualitative methods. METHODS: Semi-structured interviews were used at baseline and six months to explore carers' perception of a good therapy, the advantages and disadvantages of the different services and their fulfilment with the services. In-depth thematic analysis was carried out to explore the impact of the two different methods of service delivery on carers' quality of life. RESULTS: Day hospitals provided carers with respite opportunities, whilst domiciliary stroke teams provided carers with better educational opportunities to be involved in therapy. No qualitative difference was found in the impact that the different services had on carers' quality of life, which were influenced by factors such as the degree of disruption that caring had on their lives, the loss of a shared life and the availability of social support. Ultimately, carers saw the services as providing benefit for survivors and not themselves. CONCLUSIONS: Domiciliary stroke teams provided informal stroke carers with skills that could help improve postdischarge stroke rehabilitation amongst stroke survivors. Informal carers also benefited from the respite elements of day hospital. A mixed model using both domiciliary care and day hospital care, could provide carers with the benefits of education, convenience and respite.

A study to evaluate the met and unmet needs of members belonging to Young Stroke groups affiliated with the Stroke Association.

PURPOSE: To examine the met and unmet needs of young people with stroke who accessed services run by the Stroke Association. METHODS: One hundred and thirty-five out of 200 members of 14 participating Young Stroke groups completed and returned the postal version of the Southampton Needs Assessment Questionnaire for Stroke questionnaire. RESULTS: Young Stroke group members had a median of five unmet needs. An analysis of these needs showed that information about the person's stroke, help with finances, assistance with non-care activities (e.g. help with social activities) and help with maintaining intellectual fulfilment were the four most frequently reported ones. There was no association between the number of unmet needs reported and factors such as age, time since stroke and social class. CONCLUSIONS: Young Stroke group members may be more aware of what their unmet needs are and have better knowledge in accessing existing health and social service resources. Nevertheless, these findings show that people with stroke considered the provision of information geared to their own needs as important. Statutory services should personalize information to individual's specific situation and should also tackle other problems, not only physical ones, experienced by young people with strokes.

The unmet needs of young people who have had a stroke: results of a national UK survey.

BACKGROUND: Most stroke-related studies have consisted of people over the age of 65. This study examined the unmet needs of young people with stroke, living in community housing in the UK. METHOD: People with a stroke (>1 year ago), in two age bands (18-45; 46-65) were sent the Southampton Needs Assessment Questionnaire for people with Stroke. RESULTS: 315 out of 639 (49%) questionnaires were returned (mean age 55, SD 9; 189 males, 126 females). The median number of unmet needs reported was two (IQR 0-6). The most frequently reported unmet needs were: provision of information about the responders' stroke (45%); assistance with finances (24%); non-care activities (19%); and intellectual fulfillment (17%). Responders in the younger age group reported significantly more unmet needs than responders in the older age group (for a holiday, intellectual fulfillment and family support). Responders with poor mobility reported significantly more unmet needs than responders with average and good mobility for 15 unmet needs (three most pressing: respite care/short breaks; adaptations; and access to community environment). Responders who did not return to work reported significantly more unmet needs than responders who had reduced hours or changed jobs and people who returned to the same job with the same hours for seven unmet needs (three most pressing: help with finances; a holiday and speech therapy). CONCLUSION: People of younger age, with poorer mobility and those unable to return to work, report most unmet needs. Further work needs to be done within the community, with employers and professionals, in relation to education and the provision of specifically targeted information in order to facilitate participation and autonomy for people with stroke.

The impact of stroke on informal carers: a literature review.

Stroke incidence is set to rise in Western societies as population projections predict an increase in the proportion of older people. Most of these stroke survivors are supported by close kin members who play an important role in the rehabilitation and care of this group. The objective of this review was to establish the following: (a) to evaluate the impact of the stroke on the informal carers' quality of life, (b) to identify factors which help carers to cope with their caring role, (c) to evaluate health service provision for stroke carers. A systematic literature search using BIDS-EMBASE, MEDLINE and PSYCHLIT, identified 31 relevant studies. These showed that most studies concentrated on carers' psychological health and the negative impact that the stroke had. Carers ability to cope with the stroke was enhanced both by the use of positive coping strategies and more concrete measures e.g. more stroke information. Furthermore, whilst most carers were generally satisfied with health services, the few interventions directed at improving carer outcomes showed mixed results. The studies reviewed had many limitations; few gave definitions of 'informal' carer and there was a predominant use of cross-sectional studies and non-standardised outcome measures. Future studies should broaden their research question to evaluate quality of life, using standardised measures to do this and employing either a longitudinal or randomised control design to improve the robustness of results. More studies are also needed evaluating the effectiveness of health services on carers' quality of life.

The good and bad death perceptions of health professionals working in palliative care.

The development of palliative care originated from shortcomings in mainstream health services. Palliative care aims to cater for both the psycho-social needs of dying patients and the allieviation of their physical symptoms. This is reflected by the good and bad death perceptions of palliative care workers, though increasing signs of institutionalization in palliative care have challenged the idealization of a good death. This study aimed to investigate the health professionals' perception of both a good and a bad death and their perception of patients' awareness context. Seventy questionnaires were distributed to nurses and social workers. The 50 returned questionnaires revealed that health professionals perceived a good death as controlling the patients' physical symptoms and psychologically preparing them, whilst a bad death was perceived as the inability to control pain and deal with any psychological distress. Factor analysis identified three main factors (lack of patient distress, patient control and staff's supporting role perceptions) in the perception of a good death whereas four main factors (the negative effect of death on the family, a patient's non-acceptance of death, not dealing with patients' fears and the age of a dying person) were identified with the perception of a bad death. Overall, health professionals perceived themselves to be open and sensitive in communicating with patients although over half felt poorly supported by other staff.