Analysis of multispectral imaging with the AstroPath platform informs efficacy of PD-1 blockade

Next-generation tissue-based biomarkers for immunotherapy will likely include the simultaneous analysis of multiple cell types and their spatial interactions, as well as distinct expression patterns of immunoregulatory molecules. Here, we introduce a comprehensive platform for multispectral imaging and mapping of multiple parameters in tumor tissue sections with high-fidelity single-cell resolution. Image analysis and data handling components were drawn from the field of astronomy. Using this “AstroPath” whole-slide platform and only six markers, we identified key features in pretreatment melanoma specimens that predicted response to anti–programmed cell death-1 (PD-1)–based therapy, including CD163+PD-L1– myeloid cells and CD8+FoxP3+PD-1low/mid T cells. These features were combined to stratify long-term survival after anti–PD-1 blockade. This signature was validated in an independent cohort of patients with melanoma from a different institution.

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Spatial and Temporal Changes in PD-L1 Expression in Cancer: The Role of Genetic Drivers, Tumor Microenvironment and Resistance to Therapy

International Journal of Molecular Sciences ◽

10.3390/ijms21197139 ◽

2020 ◽

Vol 21 (19) ◽

pp. 7139 ◽

Cited By ~ 1

Author(s):

Elena Shklovskaya ◽

Helen Rizos

Keyword(s):

Tumor Microenvironment ◽

Patient Outcomes ◽

Cytotoxic T Lymphocyte ◽

Cell Types ◽

Development Of Resistance ◽

Improve Patient Selection ◽

Potential Biomarker ◽

Programmed Cell Death 1 ◽

Tumor Biopsies

Immunotherapies blocking immune inhibitory receptors programmed cell death-1 (PD-1) and cytotoxic T-lymphocyte-associated protein-4 (CTLA-4) on T-cells have dramatically improved patient outcomes in a range of advanced cancers. However, the lack of response, and the development of resistance remain major obstacles to long-term improvements in patient outcomes. There is significant interest in the clinical use of biomarkers to improve patient selection, and the expression of PD-1 ligand 1 (PD-L1) is often reported as a potential biomarker of response. However, accumulating evidence suggests that the predictive value of PD-L1 expression in tumor biopsies is relatively low due, in part, to its complex biology. In this review, we discuss the biological consequences of PD-L1 expression by various cell types within the tumor microenvironment, and the complex mechanisms that regulate PD-L1 expression at the genomic, transcriptomic and proteomic levels.

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COVID-19-related anosmia is associated with viral persistence and inflammation in human olfactory epithelium and brain infection in hamsters

Science Translational Medicine ◽

10.1126/scitranslmed.abf8396 ◽

2021 ◽

pp. eabf8396

Author(s):

Guilherme Dias de Melo ◽

Françoise Lazarini ◽

Sylvain Levallois ◽

Charlotte Hautefort ◽

Vincent Michel ◽

...

Keyword(s):

Olfactory Epithelium ◽

Cell Types ◽

Brain Infection ◽

Olfactory Neuroepithelium ◽

Optimal Medical Management ◽

Infected Cells ◽

Multiple Cell ◽

Lung Pathogenesis ◽

Taste Dysfunction

Whereas recent investigations have revealed viral, inflammatory and vascular factors involved in SARS-CoV-2 lung pathogenesis, the pathophysiology of neurological disorders in COVID-19 remains poorly understood. Olfactory and taste dysfunction are common in COVID-19, especially in mildly symptomatic patients. Here, we conducted a virologic, molecular, and cellular study of the olfactory neuroepithelium of seven patients with COVID-19 presenting with acute loss of smell. We report evidence that the olfactory neuroepithelium may be a major site of SARS-CoV2 infection with multiple cell types, including olfactory sensory neurons, support cells, and immune cells, becoming infected. SARS-CoV-2 replication in the olfactory neuroepithelium was associated with local inflammation. Furthermore, we showed that SARS-CoV-2 induced acute anosmia and ageusia in golden Syrian hamsters, lasting as long as the virus remained in the olfactory epithelium and the olfactory bulb. Finally, olfactory mucosa sampling from patients showing long-term persistence of COVID-19-associated anosmia revealed the presence of virus transcripts and of SARS-CoV-2-infected cells, together with protracted inflammation. SARS-CoV-2 persistence and associated inflammation in the olfactory neuroepithelium may account for prolonged or relapsing symptoms of COVID-19, such as loss of smell, which should be considered for optimal medical management of this disease.

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Sclerosing Hemangioma of the Lung

Archives of Pathology & Laboratory Medicine ◽

10.5858/133.5.820 ◽

2009 ◽

Vol 133 (5) ◽

pp. 820-825 ◽

Cited By ~ 8

Author(s):

C. P. T. Joren B. Keylock ◽

Jeffrey R. Galvin ◽

Teri J. Franks

Keyword(s):

Regional Lymph Node ◽

Benign Lesion ◽

Surgical Excision ◽

Cell Types ◽

Additional Treatment ◽

Term Survival ◽

Sclerosing Hemangioma ◽

Architectural Patterns ◽

Node Metastases

Abstract We present a brief review of sclerosing hemangioma, an uncommon but histologically distinctive neoplasm of the lung. Based on immunohistochemical and molecular findings, sclerosing hemangioma is thought to be derived from incompletely differentiated respiratory epithelium. Sclerosing hemangiomas typically present as asymptomatic, peripheral, solitary, well-circumscribed lesions in women with a mean age at diagnosis in the fifth decade. Rare cases are reported to have regional lymph node metastases; however, metastases do not appear to affect long-term survival. Histologically, sclerosing hemangioma is characterized by a distinct constellation of findings including 2 epithelial cell types, surface cells and round cells, which form 4 architectural patterns, papillary, sclerotic, solid, and hemorrhagic. Sclerosing hemangioma of the lung is generally considered to be a benign lesion, and surgical excision is curative without the need for additional treatment.

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Mucin mRNA Expression in Normal and Vasomotor Inferior Turbinates

American Journal of Rhinology ◽

10.2500/105065897781446685 ◽

1997 ◽

Vol 11 (4) ◽

pp. 293-302 ◽

Cited By ~ 59

Author(s):

Michelle R. Aust ◽

Cathy S. Madsen ◽

Anita Jennings ◽

Jan L. Kasperbauer ◽

Sandra J. Gendler

Keyword(s):

Respiratory Tract ◽

Expression Patterns ◽

Inferior Turbinate ◽

Cell Types ◽

Upper Respiratory Tract ◽

Vasomotor Rhinitis ◽

Disease States ◽

Multiple Cell ◽

Nasal Secretions

Mucins are the major glycoprotein component of respiratory tract secretions. Little is known about their expression in the upper respiratory tract. In order to define this expression, in situ hybridization was performed on 19 normal and 4 vasomotor rhinitis (VMR) inferior turbinates to identify mucin mRNA. MUC1, MUC2, MUC4, MUC5AC, MUC5B, and MUC7 were expressed in both the normal and VMR turbinates. MUC 4 and MUC5AC were the most highly expressed mucins. MUC1, MUC2, MUC4, and MUC5AC were expressed mainly by the epithelial border, whereas MUC5B and MUC7 were expressed by the submucosal glands. MUC1 and MUC4 exhibited a diffuse expression by multiple cell types along the mucosal border, whereas MUC2 and MUC5AC expression appeared to be limited to a subpopulation of epithelial cells, most likely goblet cells. Although MUC1, MUC4, and MUC5AC showed sporadic submucosal glandular expression, MUC5B and MUC7 appeared to be the predominant submucosal gland mucins in the inferior turbinates. MUC3 and MUC6 expression, which have been found primarily in the gastric mucosa, were not seen in any of the inferior turbinate samples examined. The only difference seen between normal and VMR turbinates was a slight decrease in MUC1 expression in the VMR group. The variety of mucins expressed and the diversity of their expression patterns may have significance in terms of the rheologic and particle clearance properties of nasal secretions. Understanding the expression patterns in normal turbinates will serve as the foundation for further study of these mucins in disease states.

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Pembrolizumab dramatically resolves choroidal metastatis from esophageal adenocarcinoma and restores vision: a case report

Oxford Medical Case Reports ◽

10.1093/omcr/omab047 ◽

2021 ◽

Vol 2021 (6) ◽

Author(s):

Dianne Barrett ◽

Andrew Sumnicht ◽

KV Chalam ◽

Micheal Rauser

Keyword(s):

Case Report ◽

Esophageal Adenocarcinoma ◽

Term Survival ◽

Gastrointestinal Malignancy ◽

Poor Vision ◽

Aggressive Cancer ◽

Programmed Cell Death 1 ◽

Ocular Metastasis

ABSTRACT Esophageal adenocarcinoma historically is an aggressive cancer with poor long-term survival. Ocular metastasis secondary to gastrointestinal malignancy is rare. In managing patients with ocular metastasis, quality of life (specifically vision preservation) is one of the most important factors patients and providers consider when deciding on a treatment regimen. Anti-programmed cell death-1 (PD-1) and PD-1 ligand (PD-L1) inhibitors such as pembrolizumab have shown promising results as second-line therapy for patient with metastatic malignancy. We describe a novel case of a functionally monocular patient with known metastatic esophageal adenocarcinoma who developed poor vision and a large choroidal lesion in his better seeing eye. The lesion regressed and vision restored to 20/20 after treatments with pembrolizumab in this case report.

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In the Absence of Apoptosis, Myeloid Cells Arrest When Deprived of Growth Factor, But Remain Viable by Consuming Extracellular Glucose

10.1101/198754 ◽

2017 ◽

Author(s):

Li Dong ◽

Boris Reljic ◽

Jen G. Cheung ◽

Elizabeth S. Ng ◽

Lisa M. Lindqvist ◽

...

Keyword(s):

Growth Factor ◽

Glucose Transporter ◽

Myeloid Cells ◽

Cell Types ◽

Term Survival ◽

Interacting Protein ◽

Extracellular Glucose ◽

Glut1 Expression ◽

High Concentrations

AbstractWithdrawal of the growth factor interleukin 3 from IL3-dependent myeloid cells causes them to undergo Bax/Bak1-dependent apoptosis, whereas factor-deprivedBax-/-Bak1-/-cells remain viable, but arrest and shrink. It was reported that withdrawal of IL3 fromBax-/-Bak1-/-cells caused decreased expression of the glucose transporter Glut1, leading to reduced glucose uptake, so that arrested cells required Atg5-dependent autophagy for long-term survival. In other cell types, a decrease in Glut1 is mediated by the thioredoxin-interacting protein Txnip, which is induced in IL3-dependent myeloid cells when growth factor is removed. We mutatedAtg5andTxnipby CRISPR/Cas9 and found that Atg5-dependent autophagy was not necessary for the long-term viability of cycling or arrestedBax-/-Bak1-/-cells, and that Txnip was not required for the decrease in Glut1 expression in response to IL3 withdrawal. Surprisingly, Atg5-deficientBax/Bak1double mutant cells survived for several weeks in medium supplemented with 10% fetal bovine serum (FBS), without high concentrations of added glucose or glutamine. When serum was withdrawn, the provision of an equivalent amount of glucose present in 10% FBS (~0.5 mM) was sufficient to support cell survival for more than a week, in the presence or absence of IL3. Thus,Bax-/-Bak1-/-myeloid cells deprived of growth factor consume extracellular glucose to maintain long-term viability, without a requirement for Atg5-dependent autophagy.

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Identification of MicroRNA–mRNA Networks in Melanoma and Their Association with PD-1 Checkpoint Blockade Outcomes

Cancers ◽

10.3390/cancers13215301 ◽

2021 ◽

Vol 13 (21) ◽

pp. 5301

Author(s):

Robert A. Szczepaniak Sloane ◽

Michael G. White ◽

Russell G. Witt ◽

Anik Banerjee ◽

Michael A. Davies ◽

...

Keyword(s):

Cancer Biology ◽

Cytotoxic T Lymphocyte ◽

Mechanisms Of Resistance ◽

Term Survival ◽

Programmed Cell Death 1 ◽

Melanoma Treatment ◽

Melanoma Patients ◽

Poor Outcomes ◽

Insight Into

Metastatic melanoma is a deadly malignancy with poor outcomes historically. Immuno-oncology (IO) agents, targeting immune checkpoint molecules such as cytotoxic T-lymphocyte associated protein-4 (CTLA-4) and programmed cell death-1 (PD-1), have revolutionized melanoma treatment and outcomes, achieving significant response rates and remarkable long-term survival. Despite these vast improvements, roughly half of melanoma patients do not achieve long-term clinical benefit from IO therapies and there is an urgent need to understand and mitigate mechanisms of resistance. MicroRNAs are key post-transcriptional regulators of gene expression that regulate many aspects of cancer biology, including immune evasion. We used network analysis to define two core microRNA–mRNA networks in melanoma tissues and cell lines corresponding to ‘MITF-low’ and ‘Keratin’ transcriptomic subsets of melanoma. We then evaluated expression of these core microRNAs in pre-PD-1-inhibitor-treated melanoma patients and observed that higher expression of miR-100-5p and miR-125b-5p were associated with significantly improved overall survival. These findings suggest that miR-100-5p and 125b-5p are potential markers of response to PD-1 inhibitors, and further evaluation of these microRNA–mRNA interactions may yield further insight into melanoma resistance to PD-1 inhibitors.

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A New Organotypic Culture of Adipose Tissue Fragments Maintains Viable Mature Adipocytes for a Long Term, Together with Development of Immature Adipocytes and Mesenchymal Stem Cell-Like Cells

Endocrinology ◽

10.1210/en.2008-0525 ◽

2008 ◽

Vol 149 (10) ◽

pp. 4794-4798 ◽

Cited By ~ 32

Author(s):

Emiko Sonoda ◽

Shigehisa Aoki ◽

Kazuyoshi Uchihashi ◽

Hidenobu Soejima ◽

Sachiko Kanaji ◽

...

Keyword(s):

Adipose Tissue ◽

Organotypic Culture ◽

Three Dimensional ◽

Collagen Gel ◽

Cell Types ◽

Ppar Γ ◽

Tnf Α ◽

Multiple Cell ◽

Related Agent

Adipose tissue that consists of mature and immature adipocytes is suggested to contain mesenchymal stem cells (MSCs), but a culture system for analyzing their cell types within the tissue has not been established. Here we show that three-dimensional collagen gel culture of rat sc adipose tissue fragments maintained viable mature adipocytes for a long term, producing immature adipocytes and MSC-like cells from the fragments, using immunohistochemistry, ELISA, and real time RT-PCR. Bromodeoxyuridine uptake of mature adipocytes was detected. Adiponectin and leptin, and adipocyte-specific genes of adiponectin, leptin, and PPAR-γ were detected in culture assembly, whereas the lipogenesis factor insulin (20 mU/ml) and inflammation-related agent TNF-α (2 nm) increased and decreased, respectively, all of their displays. Both spindle-shaped cell types with oil red O-positive lipid droplets and those with expression of MSC markers (CD105 and CD44) developed around the fragments. The data indicate that adipose tissue-organotypic culture retains unilocular structure, proliferative ability, and some functions of mature adipocytes, generating both immature adipocytes and CD105+/CD44+ MSC-like cells. This suggests that our method will open up a new way for studying both multiple cell types within adipose tissue and the cell-based mechanisms of obesity and metabolic syndrome.

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Optimized Culture Conditions for Improved Growth and Functional Differentiation of Mouse and Human Colon Organoids

Frontiers in Immunology ◽

10.3389/fimmu.2020.547102 ◽

2021 ◽

Vol 11 ◽

Author(s):

Sarah S. Wilson ◽

Martha Mayo ◽

Terry Melim ◽

Heather Knight ◽

Lori Patnaude ◽

...

Keyword(s):

Epithelial Cell ◽

Current Knowledge ◽

Human Colon ◽

Cell Types ◽

Functional Differentiation ◽

Culture Conditions ◽

Conditioned Media ◽

Term Survival ◽

The Impact

Background & AimsDiligent side-by-side comparisons of how different methodologies affect growth efficiency and quality of intestinal colonoids have not been performed leaving a gap in our current knowledge. Here, we summarize our efforts to optimize culture conditions for improved growth and functional differentiation of mouse and human colon organoids.MethodsMouse and human colon organoids were grown in four different media. Media-dependent long-term growth was measured by quantifying surviving organoids via imaging and a cell viability readout over five passages. The impact of diverse media on differentiation was assessed by quantifying the number of epithelial cell types using markers for enterocytes, stem cells, Goblet cells, and enteroendocrine cells by qPCR and histology upon removal of growth factors.ResultsIn contrast to Wnt3a-conditioned media, media supplemented with recombinant Wnt3a alone did not support long-term survival of human or mouse colon organoids. Mechanistically, this observation can be attributed to the fact that recombinant Wnt3a did not support stem cell survival or proliferation as demonstrated by decreased LGR5 and Ki67 expression. When monitoring expression of markers for epithelial cell types, the highest level of organoid differentiation was observed after combined removal of Wnt3a, Noggin, and R-spondin from Wnta3a-conditioned media cultures.ConclusionOur study defined Wnt3a-containing conditioned media as optimal for growth and survival of human and mouse organoids. Furthermore, we established that the combined removal of Wnt3a, Noggin, and R-spondin results in optimal differentiation. This study provides a step forward in optimizing conditions for intestinal organoid growth to improve standardization and reproducibility of this model platform.

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