Patient-derived organoids (PDOs) have recently emerged as robust preclinical models; however, their potential to predict clinical outcomes in patients has remained unclear. We report on a living ...biobank of PDOs from metastatic, heavily pretreated colorectal and gastroesophageal cancer patients recruited in phase 1/2 clinical trials. Phenotypic and genotypic profiling of PDOs showed a high degree of similarity to the original patient tumors. Molecular profiling of tumor organoids was matched to drug-screening results, suggesting that PDOs could complement existing approaches in defining cancer vulnerabilities and improving treatment responses. We compared responses to anticancer agents ex vivo in organoids and PDO-based orthotopic mouse tumor xenograft models with the responses of the patients in clinical trials. Our data suggest that PDOs can recapitulate patient responses in the clinic and could be implemented in personalized medicine programs.
Atherosclerosis is a lipid-driven disease of the artery characterized by chronic non-resolving inflammation. Despite availability of excellent lipid-lowering therapies, atherosclerosis remains the ...leading cause of disability and death globally. The demonstration that suppressing inflammation prevents the adverse clinical manifestations of atherosclerosis in recent clinical trials has led to heightened interest in anti-inflammatory therapies. In this review, we briefly highlight some key anti-inflammatory and pro-resolution pathways, which could be targeted to modulate pathogenesis and stall atherosclerosis progression. We also highlight key challenges that must be overcome to turn the concept of inflammation targeting therapies into clinical reality for atherosclerotic heart disease.
ATR inhibitors (ATRi) are in early phase clinical trials and have been shown to sensitize to chemotherapy and radiotherapy preclinically. Limited data have been published about the effect of these ...drugs on the tumor microenvironment.
We used an immunocompetent mouse model of HPV-driven malignancies to investigate the ATR inhibitor AZD6738 in combination with fractionated radiation (RT). Gene expression analysis and flow cytometry were performed posttherapy.
Significant radiosensitization to RT by ATRi was observed alongside a marked increase in immune cell infiltration. We identified increased numbers of CD3
and NK cells, but most of this infiltrate was composed of myeloid cells. ATRi plus radiation produced a gene expression signature matching a type I/II IFN response, with upregulation of genes playing a role in nucleic acid sensing. Increased MHC I levels were observed on tumor cells, with transcript-level data indicating increased antigen processing and presentation within the tumor. Significant modulation of cytokine gene expression (particularly CCL2, CCL5, and CXCL10) was found
, with
data indicating CCL3, CCL5, and CXCL10 are produced from tumor cells after ATRi + RT.
We show that DNA damage by ATRi and RT leads to an IFN response through activation of nucleic acid-sensing pathways. This triggers increased antigen presentation and innate immune cell infiltration. Further understanding of the effect of this combination on the immune response may allow modulation of these effects to maximize tumor control through antitumor immunity.
Behavioral technicians (BT) within the field of applied behavior analysis may be at greater risk for experiencing burnout and stress due to the nature of their clients, job demands, and work ...environments. Burnout and stress may negatively impact BT’s work performances, more specifically, their treatment integrity. Acceptance and Commitment Training (ACT) may be a useful tool to address the private events as well as the covert and overt behaviors associated with burnout and stress. The purpose of this study was to investigate the effects of an ACT intervention on improving treatment integrity and reducing work-related burnout and stress amongst BTs. Four BTs participated in an ACT workshop, and their treatment integrity as well as their burnout and stress levels were measured prior to and following the ACT workshop. Treatment integrity increased for all participants, suggesting that ACT-based interventions may be an effective approach to improving work performance (i.e., treatment integrity) amongst BTs who may experience workplace burnout and stress.
Soft tissue sarcomas (STS) are rare and diverse mesenchymal cancers with limited treatment options. Here we undertake comprehensive proteomic profiling of tumour specimens from 321 STS patients ...representing 11 histological subtypes. Within leiomyosarcomas, we identify three proteomic subtypes with distinct myogenesis and immune features, anatomical site distribution and survival outcomes. Characterisation of undifferentiated pleomorphic sarcomas and dedifferentiated liposarcomas with low infiltrating CD3 + T-lymphocyte levels nominates the complement cascade as a candidate immunotherapeutic target. Comparative analysis of proteomic and transcriptomic profiles highlights the proteomic-specific features for optimal risk stratification in angiosarcomas. Finally, we define functional signatures termed Sarcoma Proteomic Modules which transcend histological subtype classification and show that a vesicle transport protein signature is an independent prognostic factor for distant metastasis. Our study highlights the utility of proteomics for identifying molecular subgroups with implications for risk stratification and therapy selection and provides a rich resource for future sarcoma research.
Introduction: Tuberculosis (TB) is a communicable disease. The management, as well as the outcome, depends upon the early detection and diagnosis of the disease. Delay in diagnosis leads to worsening ...of the patient's condition as well as the emergence of multi-resistant bacilli. Objectives: The objectives were to know how many persons had diagnostic and treatment delays, as well as the factors that were linked to delayed care-seeking (patient delay) and diagnosis by health-care professionals (health-care system delay) among newly diagnosed pulmonary TB patients. Methodology: A cross-sectional study of 115 diagnosed pulmonary TB patients was done. The time between the initial symptoms and the first encounter with a hospital institution was referred to as the patient's delay. The time between first contact with a health-care facility and diagnosis confirmation was referred to as the diagnostic delay. A cutoff threshold of 4 weeks was used to determine the diagnostic delay. A patient delay of more than 2 weeks and a health-care system delay exceeding 2 weeks were also evaluated. Results: In this study, 94 (81.7%), 69 (60%), and 100 (87%) subjects had patient, health system, and diagnostic delays, respectively. Conclusion: The majority of the participants in the study experienced a diagnostic delay, followed by patient and health-care system delays. The educational status, income status, distance to a health care facility, cost, initial consultation with the type of health-care personnel in the health-care system, and frequency of consultations were associated with the delays.
BackgroundTriple negative breast cancer (TNBC) remains a challenge to treat due to its heterogeneous tumour microenvironment, rapid metastasis, and high recurrence rates.1 TNBCs are currently still ...treated with chemo- and radiotherapy, both of which cause significant side effects.1 Interleukin-12 (IL-12) is a powerful anti-tumour cytokine but its use in the clinic is limited by significant toxicity.2 We bioengineered IL-12 by conjugating it to both a collagen-binding domain (CBD) and a mask. The former delivers the IL-12 payload into the collagenous stroma of solid tumours,3whilst the latter temporarily suppresses IL-12 activity in the bloodstream to suppress toxicity. The mask is linked to CBD-IL12 using a protein linker, that is cleaved by proteases upon infiltrating the tumour stroma. The removal of the mask activates masked CBD-IL12 (M-CBD-IL12) in a tumour-specific manner.MethodsM-CBD-IL12 with a protease-sensitive linker was produced in HEK293F cells and purified using immobilised metal affinity and size exclusion chromatography. Murine tumour lysate and serum were used to assess the cleaving of the linker through western blotting and flow cytometry. M-CBD-IL12 was tested in two murine models of TNBC (EMT6/4T1), each with unique immune landscapes. Plasma was collected for analysis of cytokine markers and tumours were analysed using flow cytometry.ResultsM-CBD-IL12 significantly prolonged the survival of mice with orthotopic EMT6 tumours, which are considered more myeloid cell infiltrated. A complete remission was seen in 4 out of 6 mice treated with M-CBD-IL12, each of which remained tumour-free for 284 days following tumour inoculation. In an intervention setting, the IL-12- and M-CBD-IL12-treated mice had significantly smaller tumours than PBS controls. Mice treated with IL-12 or M-CBD-IL12 had significantly higher levels of intra-tumoural IFN-gamma with significantly fewer T regulatory cells. The CD8+ T-cells expressed significantly less PD-1 following M-CBD-IL12; this suggests M-CBD-IL12 reduces immunosuppressive pathways in the EMT6 tumour microenvironment. Despite similar efficacies, the mice treated with M-CBD-IL12 had significantly less plasma IFN-gamma (toxicity marker) than mice treated with IL-12, suggesting the mask and CBD are suppressing IL-12-related toxicity. The 4T1 model of TNBC is thought to be more T-cell inflamed; M-CBD-IL12 did not prolong survival of mice with 4T1 tumours, suggesting a selective efficacy of M-CBD-IL12 for TNBC.ConclusionsM-CBD-IL12 cures murine EMT6 TNBC with low toxicity and prevents its recurrence long-term. Further studies are required to understand the immunological mechanism behind the observed responses.AcknowledgementsWe would like to thank the Cancer Research UK Convergence Science Centre for funding this research and the Institute of Cancer Research Core Facilities for their support.ReferencesMarra A, Trapani D, Viale G, et al. Practical classification of triple-negative breast cancer: intratumoral heterogeneity, mechanisms of drug resistance, and novel therapies. npj Breast Cancer 2020;6:54.Nguyen KG, Vrabel MR, Mantooth SM, et al. Localized Interleukin-12 for Cancer Immunotherapy. Frontiers in Immunology 2020;11.Mansurov A, Ishihara J, Hosseinchi P, et al. Collagen-binding IL-12 enhances tumour inflammation and drives the complete remission of established immunologically cold mouse tumours. Nature Biomedical Engineering 2020;4:531–543.
Previously, we classified colorectal cancers (CRCs) into five CRCAssigner (CRCA) subtypes with different prognoses and potential treatment responses, later consolidated into four consensus molecular ...subtypes (CMS). Here we demonstrate the analytical development and validation of a custom NanoString nCounter platform-based biomarker assay (NanoCRCA) to stratify CRCs into subtypes. To reduce costs, we switched from the standard nCounter protocol to a custom modified protocol. The assay included a reduced 38-gene panel that was selected using an in-house machine-learning pipeline. We applied NanoCRCA to 413 samples from 355 CRC patients. From the fresh frozen samples (n = 237), a subset had matched microarray/RNAseq profiles (n = 47) or formalin-fixed paraffin-embedded (FFPE) samples (n = 58). We also analyzed a further 118 FFPE samples. We compared the assay results with the CMS classifier, different platforms (microarrays/RNAseq) and gene-set classifiers (38 and the original 786 genes). The standard and modified protocols showed high correlation (> 0.88) for gene expression. Technical replicates were highly correlated (> 0.96). NanoCRCA classified fresh frozen and FFPE samples into all five CRCA subtypes with consistent classification of selected matched fresh frozen/FFPE samples. We demonstrate high and significant subtype concordance across protocols (100%), gene sets (95%), platforms (87%) and with CMS subtypes (75%) when evaluated across multiple datasets. Overall, our NanoCRCA assay with further validation may facilitate prospective validation of CRC subtypes in clinical trials and beyond.
Abstract
Background: Immunotherapy has resulted in a paradigm shift in the treatment of multiple solid tumors. Still, immunotherapy in unselected pancreatic ductal adenocarcinoma (PDAC) patients has ...been disappointing. This failure is likely multifactorial and associated with a high immunosuppressive and desmoplastic tumor microenvironment (TME). This unique TME in PDAC leads to various immune escape mechanisms employed by the tumor. While this could be overcome by therapy involving IL12 (or other cytokines) alone, they often fail due to immune-related adverse effects (irAEs). On the other hand, bioengineered collagen-binding domain-linked IL-12 (CBD-IL12; T- cell stimulant that delivers IL12 to collagen-rich regions) is effective in other cancers in pre-clinical settings. Here, we hypothesise that different immune (cold/hot) and collagen content in mice determines their treatment efficacy and irAEs to CBD-IL12 or combination immunotherapy. Methods: Three (7947, 7784 and 2334) out of 12 syngeneic orthotopic models of PDAC were classified into immune “hot” (immunehot) or “cold” (immunecold) with varying collagen levels (collagenhigh or collagenlow) using cross-species statistical inference (in silico analysis). Mice were treated with vehicle or two bioengineered treatment arms – CBD-IL-12 or CBD-IL-12 + anti-PD1. Tumor growth and metastasis was assessed in an interventional trial. Transcriptome profiling was performed on harvested tumors. Results: As predicted by in silico analyses, our in vivo experiments confirmed 7947 syngeneic model as immune cold (immunecold) and resistant to selected stimulatory (GITR) and inhibitory (anti-PD1 or anti-CSF1R+anti-PD1) immunotherapies. To improve the immunotherapy in this immunecold model, we utilised the opportunity that this model has increased collagen (collagenhigh), hence, treating them with CBD-IL12 or combination with anti-PD1 therapy may provide improved treatment efficacy and reduce irAEs. Remarkably, the treatments prevented distant metastasis to the liver by significantly reducing tumor burden (p<0.05) and toxicity. The mechanistic analysis demonstrated a cascade of TME changes, including increased CD8 T cells and MHCII+ myeloid cell-based antigen presentation in the treated tumors compared to the control. This increased antigen presentation was associated with macrophage repolarisation from M2 to M1. This was not the case in two other immunecold/collagenlow or immunehot/collagenhigh models. Conclusion: Our integrated strategy to pre-select mouse models based on their TME (immune/collagen) profiles improves personalised bioengineered immunotherapy response in immune cold PDAC. Furthermore, the added benefit of preventing metastasis is rather promising and warrants further investigation in aggressive disease like PDAC.
Citation Format: Chanthirika Ragulan, Patrick Varun Lawrence, Hari Ps, Krisha Desai, Jun Ishihara, Anguraj Sadanandam. Heating up immune cold pancreatic adenocarcinoma with bioengineered immunotherapy remodels tumor microenvironment and prevents metastasis in vivo abstract. In: Proceedings of the AACR Virtual Special Conference on Pancreatic Cancer; 2021 Sep 29-30. Philadelphia (PA): AACR; Cancer Res 2021;81(22 Suppl):Abstract nr PO-020.
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