EPCO-15. SPATIAL-TRANSCRIPTOMICS REVEALS UNIQUE MOLECULAR FEATURES OF FLUORESCENCE-SORTED 5-AMINOLEVULINIC ACID+ INFILTRATIVE TUMOR CELLS ASSOCIATED WITH RECURRENCE AND POOR SURVIVAL IN GLIOBLASTOMA

2021 ◽  
Vol 23 (Supplement_6) ◽  
pp. vi4-vi5
Author(s):  
Geoffroy Andrieux ◽  
Tonmoy Das ◽  
Stuart Smith ◽  
Ruman Rahman ◽  
Sajib Chakraborty
Keyword(s):  
Tumor Cells ◽  
Aminolevulinic Acid ◽  
Molecular Subtype ◽  
Wound Response ◽  
Neural Cells ◽  
Molecular Signatures ◽  
Poor Survival ◽  
Neoplastic Cells ◽  
Invasive Margin ◽  
Molecular Features

Abstract Spatiotemporal heterogeneity of glioblastoma (GBM) originating from genomic and transcriptional variation in spatially distinct sites, may contribute to subtype switching in GBM prior to and upon recurrence. Fluorescence-guided neurosurgical resection utilizing 5-aminolevulinic acid (5ALA) has enabled the isolation of infiltrative margin tumor cells (5ALA+ cells) from a background of non-neoplastic cells. To falsify the hypothesis that the 5ALA+ subpopulation(s) is defined by a unique cellular state, we have explored the spatial-transcriptomic (ST) landscape to interrogate molecular signatures unique to infiltrating 5ALA+ cells in comparison to GBM core, rim, and invasive margin non-neoplastic cells. ST analysis reveals that GBM molecular subtype plasticity is not restricted to recurrence, but manifests regionally in a cell-type-specific manner. Whilst GBM core and rim are highly enriched with Classical and Proneural subtypes, 5ALA+ cells are uniquely enriched with the Mesenchymal subtype (MES). Upregulation of the wound response pathway in 5ALA+ cells implicate hijacking of the wound healing pathway of neural cells to promote tumor growth. Exon-intron split analysis revealed an upregulation of exonic counts for MES and wound-response genes in 5ALA+ cells, implying that these genes are under active post-transcriptional control. Network analysis suggests that wound response genes, including chemokine CCL2 that recruits regulatory T-cells and monocytic myeloid-derived suppressor cells, are controlled by an IRF8-mediated transcriptional program in 5ALA+ cells. A higher stemness signature both in 5ALA+ cells and non-neoplastic cells of the invasive margin emphasizes the role of this microenvironment in stemness acquisition and defines 5ALA+ cells as a rare sub-population of GBM stem cells. Finally, we establish a link between the unique molecular-signatures of 5ALA+ cells, and poor survival and GBM recurrence. Characterization of the 5ALA+ infiltrative sub-population offers an opportunity to develop more effective GBM treatments and urges focus away from the GBM proliferative core, upon which failed targeted therapies have been predicated.

scholarly journals Spatial-transcriptomics reveals unique defining molecular features of fluorescence-sorted 5-aminolevulinic acid+ infiltrative tumor cells associated with glioblastoma recurrence and poor survival

2021 ◽  
Author(s):  
Geoffroy Andrieux ◽  
Tonmoy Das ◽  
Michaela Griffin ◽  
Stuart J. Smith ◽  
Ruman Rahman ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Aminolevulinic Acid ◽  
Molecular Subtype ◽  
Wound Response ◽  
Neural Cells ◽  
Molecular Signatures ◽  
Poor Survival ◽  
Neoplastic Cells ◽  
Invasive Margin ◽  
Molecular Features

Spatiotemporal-heterogeneity of glioblastoma (GBM) originating from the genomic and transcriptional variation in spatially distinct intra-tumor sites, may contribute to subtype switching in GBM prior to and upon recurrence. Fluorescence-guided neurosurgical resection utilizing 5-aminolevulinic acid (5ALA) has enabled the isolation of infiltrative margin tumor cells (5ALA+ cells) from a background of non-neoplastic cells. We have explored the spatial-transcriptomic (ST) landscape to interrogate molecular signatures unique to infiltrating 5ALA+ cells in comparison to GBM core, rim, and invasive margin non-neoplastic cells. ST analysis reveals that GBM molecular subtype plasticity is not restricted to recurrence, but manifests regionally in a cell-type-specific manner. Whilst GBM core and rim are highly enriched with Classical and Proneural subtypes, the unique enrichment of the Mesenchymal subtype (MES) in 5ALA+ cells supports the hypothesis that MES 5ALA+ cells may drive GBM recurrence. Upregulation of the wound response pathway in 5ALA+ cells signifies the possibility of hijacking the wound healing pathway of neural cells to promote tumor growth. Exon-intron split analysis revealed an upregulation of exonic counts for MES and wound-response genes in 5ALA+ cells, implying that these genes are under active post-transcriptional control. Network analysis suggests that wound response genes, including chemokine CCL2 that recruits regulatory T-cells and monocytic myeloid-derived suppressor cells, are controlled by an IRF8-mediated transcriptional program in 5ALA+ cells. A higher stemness signature both in 5ALA+ cells and non-neoplastic cells of the invasive margin emphasizes the role of this microenvironment in stemness acquisition and defines 5ALA+ cells as a rare sub-population of GBM stem cells. Finally, we establish a link between the unique molecular signatures of 5ALA+ cells and poor survival and GBM recurrence. Characterization of the 5ALA+ infiltrative sub-population offers an opportunity to develop more effective GBM treatments and urges focus away from the GBM proliferative core, upon which failed targeted therapies have been predicated.

scholarly journals Spatial-transcriptomics reveals unique defining molecular features of 5-aminolevulinic acid+ infiltrative tumor cells associated with glioblastoma recurrence and poor survival

2021 ◽  
Author(s):  
Geoffroy Andrieux ◽  
Tonmoy Das ◽  
Michaela Griffin ◽  
Melanie Boerries ◽  
Stuart Smith ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Aminolevulinic Acid ◽  
Residual Disease ◽  
Molecular Subtype ◽  
Brain Parenchyma ◽  
Wound Response ◽  
Poor Survival ◽  
Molecular Features ◽  
Promote Tumor Growth ◽  
Recurrent Gbm

Abstract Spatiotemporal-heterogeneity originating from genomic and transcriptional variation contributes to subtype switching in GBM prior to and upon recurrence. Fluorescence-guided neurosurgical resection utilizing 5-aminolevulinic acid (5ALA) enables the isolation of infiltrative margin tumor cells (5ALA+) from a non-neoplastic background within neighboring brain parenchyma. Spatial transcriptomics identifies GBM molecular subtype plasticity as not restricted to recurrence, but manifesting regionally in a cell-type-specific manner, where a 5ALA+ Mesenchymal subtype may drive recurrence. Exon-intron split analysis reveals hijacking of the neural wound response pathway to promote tumor growth, via IRF8-mediated post-transcriptonal control. A unique stemness index further defines 5ALA+ cells as a rare sub-population of infiltrative GBM stem cells. Finally, we establish that enriched gene signatures of 5ALA+ cells are associated with poor survival and recurrence in GBM, signifying that transition from primary to recurrent GBM is not discreet, but rather a continuum whereby 5ALA+ residual disease more closely resembles the eventual recurrent GBM.

scholarly journals Macrophages and microglia: the cerberus of glioblastoma

2021 ◽  
Vol 9 (1) ◽  
Author(s):  
Alice Buonfiglioli ◽  
Dolores Hambardzumyan
Keyword(s):  
Tumor Cells ◽  
Innate Immune System ◽  
Tumor Heterogeneity ◽  
Expression Profiles ◽  
Brain Parenchyma ◽  
Innate Immune ◽  
Malignant Growth ◽  
Neoplastic Cells ◽  
Functional Roles ◽  
The Brain

AbstractGlioblastoma (GBM) is the most aggressive and deadliest of the primary brain tumors, characterized by malignant growth, invasion into the brain parenchyma, and resistance to therapy. GBM is a heterogeneous disease characterized by high degrees of both inter- and intra-tumor heterogeneity. Another layer of complexity arises from the unique brain microenvironment in which GBM develops and grows. The GBM microenvironment consists of neoplastic and non-neoplastic cells. The most abundant non-neoplastic cells are those of the innate immune system, called tumor-associated macrophages (TAMs). TAMs constitute up to 40% of the tumor mass and consist of both brain-resident microglia and bone marrow-derived myeloid cells from the periphery. Although genetically stable, TAMs can change their expression profiles based upon the signals that they receive from tumor cells; therefore, heterogeneity in GBM creates heterogeneity in TAMs. By interacting with tumor cells and with the other non-neoplastic cells in the tumor microenvironment, TAMs promote tumor progression. Here, we review the origin, heterogeneity, and functional roles of TAMs. In addition, we discuss the prospects of therapeutically targeting TAMs alone or in combination with standard or newly-emerging GBM targeting therapies.

Unusual childhood extramedullary hematologic malignancy with natural killer cell properties that contains tropomyosin 4–anaplastic lymphoma kinase gene fusion

Blood ◽  
2001 ◽  
Vol 98 (4) ◽  
pp. 1209-1216 ◽  
Author(s):  
Sandra J. Meech ◽  
Loris McGavran ◽  
Lorrie F. Odom ◽  
Xiayuan Liang ◽  
Lynne Meltesen ◽  
...  
Keyword(s):  
Nk Cells ◽  
Natural Killer ◽  
Tumor Cells ◽  
Functional Properties ◽  
Anaplastic Lymphoma Kinase ◽  
Clinical Presentation ◽  
Hematologic Malignancy ◽  
Malignant Cells ◽  
Neoplastic Cells ◽  
Anaplastic Lymphoma

This report describes an unusual extramedullary hematologic malignancy in an 18-month-old child who presented with a capillary leak syndrome that evolved into hyperleukocytosis with malignant cells. The circulating tumor cells did not express an antigen profile typical of any subtype of leukemia commonly observed in children. Tumor cells were CD3−/CD56+; had germline TCRgenes; and strongly expressed CD30, epithelial membrane antigen, and anaplastic lymphoma kinase (ALK) consistent with a null cell anaplastic large cell lymphoma (ALCL). The malignant cells contained a t(2;19)(p23;p13.1) that interrupted ALK and translocated it to the der(19). Reverse transcriptase-polymerase chain reaction and nucleotide sequence analysis revealed fusion of ALK to tropomyosin 4, an ALK fusion partner not described previously in hematologic malignancies. The clinical presentation and phenotypic features of this malignancy were not typical for ALCL because tumor cells expressed both myeloid (CD13, CD33, HLA-DR) and natural killer (NK) cell antigens. The neoplastic cells most resembled NK cells because in addition to being CD3−/CD56+ with germline TCR genes, these cells were CD25+/CD122+/granzyme B+ and possessed the functional properties of immature NK cells. The unusual clinical presentation, immunophenotype, and functional properties of these neoplastic cells suggest that this malignancy may be derived from the putative myeloid-NK precursor cell. Furthermore co-expression of NK and ALCL features supports the concept that a minority of null-ALCL may be derived from NK cells and expands the spectrum of phenotypes that can be seen in tumors produced by ALK fusion proteins.

scholarly journals Intravascular neoplastic cells in canine cutaneous plasmacytomas

2018 ◽  
Vol 30 (2) ◽  
pp. 329-332 ◽  
Author(s):  
Gordon Ehrensing ◽  
Linden E. Craig
Keyword(s):  
Endothelial Cells ◽  
Local Recurrence ◽  
Light Microscopy ◽  
Tumor Cells ◽  
Prognostic Significance ◽  
Neoplastic Cells ◽  
Surgical Biopsy ◽  
University Of Tennessee ◽  
The University

We evaluated 134 cutaneous plasmacytomas in 125 dogs submitted to the University of Tennessee surgical biopsy service between 2009 and 2012 to determine whether the presence of intravascular neoplastic cells had prognostic significance. Tumors occurred in middle-aged to geriatric dogs (range: 5–16 y, mean: 9.6 y) and most frequently involved the skin of the head and distal limbs. Diagnoses were made based on light microscopy, and in some cases confirmed by immunoreactivity of neoplastic cells for MUM1. Tumors were categorized as having or not having intravascular neoplastic cells within sections examined. The intravascular location of tumor cells was confirmed by immunoreactivity of endothelial cells for factor VIII–related antigen in 3 cases. Neoplastic cells within vessel lumens were identified in 20 of 125 dogs (16%). Submitting veterinary practices were contacted for follow-up data on patients including local recurrence and cutaneous plasmacytomas in other locations. Follow-up information was acquired on 99 dogs (79%). Recurrence was documented in one dog with cutaneous plasmacytomas; both masses had incomplete margins and intravascular neoplastic cells. Additional distant cutaneous plasmacytomas were later diagnosed in 3 patients; none of these dogs had intravascular neoplastic cells. In no cases were cutaneous plasmacytomas suspected to be a cause of death or reason for euthanasia. Intravascular neoplastic cells were more common in tumors of the distal limbs (36%) compared to other locations (11%; p = 0.0007). The presence of intravascular neoplastic cells did not affect prognosis in cutaneous plasmacytomas.

EXTH-49. FOCUSED ULTRASOUND AND 5-ALA MEDIATED ELIMINATION OF DIFFUSE MIDLINE GLIOMA

2021 ◽  
Vol 23 (Supplement_6) ◽  
pp. vi174-vi174
Author(s):  
Luana Schaab ◽  
Yann Ferry ◽  
Mehmet Ozdas ◽  
Bettina Kritzer ◽  
Sulayman Mourabit ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Focused Ultrasound ◽  
Aminolevulinic Acid ◽  
Clinical Investigation ◽  
Protoporphyrin Ix ◽  
Brain Regions ◽  
Sonodynamic Therapy ◽  
Diffuse Infiltration ◽  
Rat Glioma ◽  
Diffuse Midline Glioma

Abstract Diffuse midline glioma (DMG) is a devastating and incurable childhood brain cancer. With a median survival of only 9 to 11 months, over 90% of children affected by DMG die within two years of diagnosis. Despite decades of research and a growing understanding of the biology of these tumors, there have been no advancements in therapies for DMGs. Tumor heterogeneity and diffuse infiltration in inoperable brain regions make these tumors uniquely difficult to manage both surgically and pharmacologically. Therefore, there is an urgent need for the exploration of novel treatment regimens. Focused Ultrasound (FUS) is an emerging technology with significant clinical potentials. Sonodynamic therapy (SDT) is an up-and-coming treatment strategy aiming to non-invasively eliminate tumor cells by acting through compounds known as sonosensitizers, which render tumor cells sensitive to ultrasound energy. Recently, 5-Aminolevulinic acid (5-ALA), an FDA-approved molecule, has been proposed as a sono-sensitizing agent. 5-ALA mediated SDT prolonged survival in C6 rat glioma models by selective elimination of tumor cells upon sonication. Mechanistically, it is thought that 5-ALA uptake and metabolic conversion into Protoporphyrin IX (PpIX) occurs preferentially in tumor cells due to differential activity of enzymes involved in heme metabolism. Here, we investigated SDT in DMG cells treated with 5-ALA. PpIX fluorescence increased linearly up to 24 h upon 5-ALA treatment and accumulated significantly more (1.6-fold, p < 0.01) when compared to C6 cells. Consequently, FUS sonication of 5-ALA treated DMG cells at 250 kHz significantly (p < 0.05) decreased DMG cell viability compared to treatment with 5-ALA or FUS alone. Here, we show the first 5-ALA mediated sonodynamic effect in DMG cells, leading to enhanced cell death. Our findings provide a rationale for considering clinical investigation of 5-ALA mediated sonodynamic therapy in DMG.

scholarly journals Cerebrospinal fluid cytomorphologic findings in 41 intracranial tumors: a retrospective review

1995 ◽  
Vol 53 (2) ◽  
pp. 218-226 ◽  
Author(s):  
Maria José Sá ◽  
Rui Vaz ◽  
Celso Cruz
Keyword(s):  
Cerebrospinal Fluid ◽  
Cell Count ◽  
Tumor Cells ◽  
Retrospective Review ◽  
Ct Scanning ◽  
Meningeal Carcinomatosis ◽  
Cerebral Metastasis ◽  
Unexpected Finding ◽  
Intracranial Tumors ◽  
Neoplastic Cells

The main objective of this retrospective review of clinical and cerebrospinal fluid (CSF) data from 41 patients with intracranial tumors diagnosed between 1975 and 1989, is to report the role that the finding of neoplastic cells in CSF plays, specially when cerebral CT-scanning and MRI were not currently done. Another objective is to study the CSF proteic abnormalities in cerebral tumors. CSF cell count, cytomorphologic pictures obtained after sedimentation and protein findings are described. Tumor cells were seen in 12 cases (29%): medulloblastomas - 6, meningeal carcinomatosis - 3, multiforme glioblastoma - 1, ependymoma -1, cerebral metastasis -1; in two cases it was an unexpected finding. We noticed that tumoral localization next to the ventricles favoured cell exfoliation. Although pleocytosis was rare and uncorrelated with the presence of neoplastic cells, pathological cytomorphologic pictures appeared in most of the cases including all "positive" ones. Our results stress that the appearance of neoplastic cells in CSF remains helpful specially when it is an unexpected finding.

scholarly journals Photoradiation models for the clinical ex vivo treatment of autologous bone marrow grafts

Blood ◽  
1987 ◽  
Vol 70 (2) ◽  
pp. 484-489
Author(s):  
J Atzpodien ◽  
SC Gulati ◽  
A Strife ◽  
BD Clarkson
Keyword(s):  
Bone Marrow ◽  
Tumor Cells ◽  
White Light ◽  
Ex Vivo ◽  
Light Exposure ◽  
B Cell Lymphoma ◽  
Autologous Bone Marrow ◽  
Neoplastic Cells ◽  
Drug Concentrations ◽  
Autologous Bone

To assess the potential of photoradiation therapy for the in vitro purging of residual tumor cells from autologous bone marrow (BM) transplants, we studied normal marrow and tumor cell clonogenicity in response to different light-activated compounds by using the fluorescent dyes dihematoporphyrin ether (DHE) and merocyanine-540 (MC- 540). After photoradiation of cells with white light, both DHE and MC- 540 showed high cytocidal activity toward lymphoid and myeloid neoplastic cells but had a significantly lesser effect on normal granulocyte-macrophage (CFU-GM), erythroid (BFU-E), and mixed colony- forming (CFU-GEMM) progenitor cells. Acute promyelocytic leukemia (HL- 60), non-B, non-T, CALLA-positive acute lymphoblastic leukemia (Reh), and diffuse histocytic B cell lymphoma (SK-DHL-2) cell lines were exposed to different drug concentrations in combination with white light at a constant illumination rate of 50,000 lux. With DHE doses varying from 2.0 to 2.5 micrograms/mL and MC-540 concentrations of 15 to 20 micrograms/mL, clonogenic tumor cells could be reduced by more than 4 logs when treated alone or in mixtures with normal irradiated human marrow cells. However, preferential cytotoxicity towards neoplastic cells was highly dependent on the mode of light activation. MC-540 had no substantial effect on malignant lymphoid (SK-DHL-2) and myeloid (HL-60) cells and on normal marrow myeloid (CFU-GM) precursors when drug incubation was performed in the dark and followed by light exposure of washed cells. Equal doses of MC-540 (15 to 20 micrograms/mL) could preferentially eliminate tumor cells under conditions of simultaneous light and drug treatment (30 minutes at 37 degrees C). When using DHE (2.5 micrograms/mL), 29.3%, 46.8%, and 27.5% of normal marrow CFU-GM, BFU-E, and CFU-GEMM, respectively, were spared after sequential drug and light exposure of cells, whereas simultaneous treatment reduced both normal (CFU-GM) and neoplastic cells below the limits of detection. In summary, our results indicate the usefulness of various photoradiation models for the ex vivo treatment of leukemic and lymphomatous bone marrow autografts.

scholarly journals Integrative analysis identifies key molecular signatures underlying neurodevelopmental deficits in fragile X syndrome

2019 ◽  
Author(s):  
Kagistia Hana Utami ◽  
Niels H. Skotte ◽  
Ana R. Colaço ◽  
Nur Amirah Binte Mohammad Yusof ◽  
Bernice Sim ◽  
...  
Keyword(s):  
Fragile X Syndrome ◽  
Fragile X ◽  
Neurodevelopmental Disorder ◽  
Embryonic Stem ◽  
Neural Progenitor Cell ◽  
Integrative Analysis ◽  
Network Activity ◽  
Neural Cells ◽  
Molecular Signatures ◽  
Neurodevelopmental Abnormalities

AbstractFragile X syndrome (FXS) is an incurable neurodevelopmental disorder with no effective treatment. FXS is caused by epigenetic silencing ofFMR1and loss of FMRP expression. To investigate the consequences of FMRP deficiency in the context of human physiology, we established isogenicFMR1knockout (FMR1KO) human embryonic stem cells (hESCs). Integrative analysis of the transcriptomic and proteomic profiles of hESC-derived FMRP-deficient neurons revealed several dysregulated pathways important for brain development including processes related to axon development, neurotransmission, and the cell cycle. We functionally validated alterations in a number of these pathways, showing abnormal neural rosette formation and increased neural progenitor cell proliferation inFMR1KO cells. We further demonstrated neurite outgrowth and branching deficits along with impaired electrophysiological network activity in FMRP-deficient neurons. Using isogenicFMR1KO hESC-derived neurons, we reveal key molecular signatures and neurodevelopmental abnormalities arising from loss of FMRP. We anticipate that theFMR1KO hESCs and the neuronal transcriptome and proteome datasets will provide a platform to delineate the pathophysiology of FXS in human neural cells.

scholarly journals PD-L1 Expression Patterns in Microsatellite Instability-High Intestinal Adenocarcinoma Subtypes

2019 ◽  
Vol 152 (3) ◽  
pp. 384-391 ◽  
Author(s):  
Jordan Roberts ◽  
Safia N Salaria ◽  
Justin Cates ◽  
Yang Wang ◽  
Cindy Vnencak-Jones ◽  
...  
Keyword(s):  
Microsatellite Instability ◽  
Tumor Cells ◽  
Mucinous Adenocarcinoma ◽  
Expression Patterns ◽  
Inflammatory Cells ◽  
Medullary Carcinoma ◽  
Molecular Features ◽  
Programmed Death ◽  
Programmed Death Protein 1 ◽  
Histologic Subtypes

Abstract Objectives To investigate patterns of programmed death protein-1 (PD-L1) expression in microsatellite instability (MSI)-high intestinal carcinomas and correlate them with pathologic and molecular features. Methods One hundred and fifteen MSI-high and 41 microsatellite stable carcinomas were included. Tumor sections were immunohistochemically labeled for PD-L1. The results were correlated with histologic subtypes, MSI, and BRAF status. Results As expected, MSI status was associated with PD-L1 expression. Among 115 MSI-high tumors, PD-L1 expression was observed on tumor cells in 28 tumors and on tumor-associated inflammatory cells in 77 tumors. Medullary carcinoma demonstrated more frequent PD-L1 expression on tumor cells than mucinous and typical adenocarcinoma. PD-L1 expression was more frequent in medullary and typical adenocarcinoma than in mucinous adenocarcinoma based on combined positive scores. Tumors with more nucleotide shifts by PCR-based MSI testing were more likely to express PD-L1. Conclusions Expression of PD-L1 is different among different histologic subtypes of MSI-high intestinal carcinomas.

Sign in / Sign up

Export Citation Format

Share Document