3D DESI-MS lipid imaging in a xenograft model of glioblastoma: a proof of principle

Abstract Desorption electrospray ionisation mass spectrometry (DESI-MS) can image hundreds of molecules in a 2D tissue section, making it an ideal tool for mapping tumour heterogeneity. Tumour lipid metabolism has gained increasing attention over the past decade; and here, lipid heterogeneity has been visualised in a glioblastoma xenograft tumour using 3D DESI-MS imaging. The use of an automatic slide loader automates 3D imaging for high sample-throughput. Glioblastomas are highly aggressive primary brain tumours, which display heterogeneous characteristics and are resistant to chemotherapy and radiotherapy. It is therefore important to understand biochemical contributions to their heterogeneity, which may be contributing to treatment resistance. Adjacent sections to those used for DESI-MS imaging were used for H&E staining and immunofluorescence to identify different histological regions, and areas of hypoxia. Comparing DESI-MS imaging with biological staining allowed association of different lipid species with hypoxic and viable tissue within the tumour, and hence mapping of molecularly different tumour regions in 3D space. This work highlights that lipids are playing an important role in the heterogeneity of this xenograft tumour model, and DESI-MS imaging can be used for lipid 3D imaging in an automated fashion to reveal heterogeneity, which is not apparent in H&E stains alone.

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DDRE-50. INVESTIGATING THE ROLE OF LonP1 IN GLIOBLASTOMA TUMOR PROGRESSION

Neuro-Oncology ◽

10.1093/neuonc/noab196.334 ◽

2021 ◽

Vol 23 (Supplement_6) ◽

pp. vi85-vi85

Author(s):

Christopher Douglas ◽

Daniela Bota ◽

Kaijun Di ◽

Bhaskar Das ◽

Javier Lepe

Keyword(s):

Tumor Progression ◽

Malignant Gliomas ◽

Xenograft Model ◽

Treatment Resistance ◽

Proteasome Inhibition ◽

High Rate ◽

Who Grade ◽

Dismal Prognosis ◽

Stem Cell Lines ◽

Novel Strategy

Abstract Glioblastoma (GBM), a WHO grade IV brain cancer, exhibits strong treatment resistance and a high rate of reoccurrence, which gives it a dismal prognosis, a 5% survival rate in the first 5 years. LonP1, a mitochondrial master regulator, can drive metabolic transformation, cytokine production, EMT, and treatment resistance in various cancer types, but its role in GBM remains unexplored. Our research group has previously shown that LonP1 is overexpressed in human malignant gliomas, particularly glioblastoma, and that this is associated with disease prognosis. Here, we present findings that demonstrate that LonP1 seems to drive enhanced tumor progression, invasiveness, angiogenesis in different high grade glioblastomas based on TCGA-subtype. Furthermore, in collaboration with Professor Bhaskar Das, we have validated a lead compound, BT317, with on-target inhibition of LonP1 protease activity. BT317 has enhanced activity against glioma stem cell lines (GSC) and has demonstrated low toxicity and efficacy in an intracranial xenograft model. This preliminary data highlights the potential of using combinatorial, pharmacological LonP1 and proteasome inhibition as a novel strategy for targeting specific subtypes of GBM.

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Lipid dynamics in LPS-induced neuroinflammation by DESI-MS imaging

Brain Behavior and Immunity ◽

10.1016/j.bbi.2019.01.029 ◽

2019 ◽

Vol 79 ◽

pp. 186-194 ◽

Cited By ~ 3

Author(s):

Onésia Cristina Oliveira-Lima ◽

Juliana Carvalho-Tavares ◽

Marcella F. Rodrigues ◽

Marcus V. Gomez ◽

A.C.P. Oliveira ◽

...

Keyword(s):

Lipid Dynamics ◽

Ms Imaging ◽

Desi Ms

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Abstract PO-042: A multimodal analysis in breast cancer: Revealing metabolic heterogeneity using DESI-MS imaging with Laser-microdissection coupled transcriptome approach

10.1158/1538-7445.tumhet2020-po-042 ◽

2020 ◽

Author(s):

Emine Kazanc ◽

Evi Karali ◽

Vincen Wu ◽

Paolo Inglese ◽

James McKenzie ◽

...

Keyword(s):

Breast Cancer ◽

Laser Microdissection ◽

Multimodal Analysis ◽

Ms Imaging ◽

Desi Ms ◽

Metabolic Heterogeneity

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187 OVARIAN CYCLE LIPID DYNAMICS REVEALED BY DESI-MS IMAGING AND MORPHOLOGICALLY-DRIVEN MULTIVARIATE STATISTICS

Reproduction Fertility and Development ◽

10.1071/rdv27n1ab187 ◽

2015 ◽

Vol 27 (1) ◽

pp. 184

Author(s):

A. K. Jarmusch ◽

C. R. Ferreira ◽

L. S. Eberlin ◽

V. Pirro

Keyword(s):

Multivariate Statistics ◽

Ovarian Tissue ◽

Univariate Analysis ◽

Ambient Ionization ◽

Negative Ion ◽

Corpora Lutea ◽

Ionization Mass ◽

Ms Imaging ◽

Adrenic Acid ◽

Desi Ms

Understanding the role of lipid metabolism in ovarian physiology is crucial for the progression of reproductive biotechnology. The aim in this work was to explore the lipid composition and dynamics of ovarian tissue, specifically the stroma, follicles, and corpora lutea. Desorption electrospray ionization–mass spectrometry (DESI-MS), an ambient ionization technique, was applied in this investigation, acquiring chemical and spatial information simultaneously. A morphologically-friendly solvent, dimethylformamide-acetonitrile (1 : 1), was used for DESI-MS imaging which allowed for ovarian lipid characterisation and subsequent staining (hematoxylin and eosin) providing morphological information. By this approach, regions-of-interest (ROI) were selected from bovine (n = 8), swine (n = 3), and mice (n = 5) ovaries (including pre-pubescent and cycling adults) based on the stained morphological structures. ROI for stroma (n = 54), follicles (n = 89), and corpora lutea (n = 61) were selected and chemically profiled. Tissue sections (20 μm) were thaw mounted onto glass microscope slides and stored at –80°C until analysis. A linear ion trap mass spectrometer equipped with a custom DESI-MS imaging stage was operated in the negative ion mode (m/z 200 to 1000). A 300 × 300 µm pixel size was used in DESI-MS imaging of ovarian tissue. Hyperspectral DESI images were reconstructed and processed by principal component analysis (PCA) that allowed visualisation of relationships among spatial (i.e. morphology) and chemical features. Ions indicated by PCA were analysed using univariate analysis (ANOVA), supporting the significance of particular lipids between morphological structures, e.g. adrenic acid (P = 1.7 × 10–8) and m/z 836 (P = 8.9 × 10–9) between corpora lutea and follicles. All morphological structures could be differentiated by multivariate statistics (>90% prediction rate) independent of the species, indicating conserved lipid constitution. Smaller differences in the lipid profiles were noted between species, poly-ovulatory and mono-ovulatory species, and reproductive maturation. A large variety and abundance of lipids was observed in corpora lutea and follicles, where steroidogenesis is a prominent physiological activity. Additional insight into ovarian physiology was gained with the detection of arachidonic and adrenic acid. The spatial relationship of arachidonic and adrenic acid with the corpora lutea – the former is a known prostaglandin precursor and key signalling molecule in steroidogenesis regulation and the latter is metabolized in the prostaglandin pathway by the same enzymes – suggests the latter may also have a role in steroidogenesis regulation, previously unseen in ovarian physiology. DESI-MS imaging with morphologically-driven statistical analysis proved efficient in relating and interpreting the chemical and morphological features. This methodology can by further applied to unravel complex ovarian-related physiological mechanisms and to other physiological and physiopathological models.

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Cerebral Lipid Dynamics in Chronic Cerebral Hypoperfusion Model by DESI-MS Imaging

Neuroscience ◽

10.1016/j.neuroscience.2019.11.014 ◽

2020 ◽

Vol 426 ◽

pp. 1-12 ◽

Cited By ~ 2

Author(s):

Dryelle L.R. Severiano ◽

Onésia C. Oliveira-Lima ◽

Géssica A. Vasconcelos ◽

Bruno Lemes Marques ◽

Gustavo Almeida de Carvalho ◽

...

Keyword(s):

Chronic Cerebral Hypoperfusion ◽

Cerebral Hypoperfusion ◽

Lipid Dynamics ◽

Ms Imaging ◽

Desi Ms

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Understanding the Molecular Signatures in Leaves and Flowers by Desorption Electrospray Ionization Mass Spectrometry (DESI MS) Imaging

Journal of Agricultural and Food Chemistry ◽

10.1021/jf4011998 ◽

2013 ◽

Vol 61 (31) ◽

pp. 7477-7487 ◽

Cited By ~ 43

Author(s):

R. G. Hemalatha ◽

T. Pradeep

Keyword(s):

Mass Spectrometry ◽

Electrospray Ionization ◽

Electrospray Ionization Mass Spectrometry ◽

Desorption Electrospray Ionization ◽

Ionization Mass Spectrometry ◽

Ionization Mass ◽

Molecular Signatures ◽

Ms Imaging ◽

Desi Ms

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TCF4 induces enzalutamide resistance via neuroendocrine differentiation in prostate cancer

10.1101/560821 ◽

2019 ◽

Author(s):

Geun Taek Lee ◽

Won Tae Kim ◽

Young Suk Kwon ◽

Ganesh Palapattu ◽

Rohit Mehra ◽

...

Keyword(s):

Prostate Cancer ◽

Cell Line ◽

Xenograft Model ◽

Neuroendocrine Differentiation ◽

Treatment Resistance ◽

Parental Cell ◽

Parental Cell Line ◽

Castration Resistant Prostate Cancer ◽

Expression Levels ◽

Mouse Xenograft Model

AbstractIn treating patients with castration resistant prostate cancer (CRPC), enzalutamide, the second-generation androgen receptor (AR) antagonist, is an accepted standard of care. However, clinical benefits are limited to a median time of 4.8 months because resistance inevitably emerges. To determine the mechanism of treatment resistance, we carried out a RNA sequence analysis and found increased expression levels of neuroendocrine markers in the enzalutamide-resistant LNCaP human prostate cancer (CaP) cell line when compared to the parental cell line. Subsequent studies demonstrated that TCF4, a transcription factor implicated in Wnt signaling, mediated neuroendocrine differentiation (NED) in response to enzalutamide treatment and was elevated in the enzalutamide-resistant LNCaP. In addition, we observed that PTHrP mediated enzalutamide resistance in tissue culture and inducible TCF4 overexpression resulted in enzalutamide-resistance in a mouse xenograft model. Finally, small molecule inhibitors of TCF4 or PTHrP partially reversed enzalutamide resistance in CaP cells. When tissues obtained from men who died of metastatic CaP were examined, a positive correlation was found between the expression levels of TCF4 and PTHrP. Taken together, the current results indicate that TCF4 induces enzalutamide resistance via NED in CaP.

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A Study on the Practicalities of Using Volumetric Three-Dimensional Imaging Devices in Design

Volume 3: 26th Computers and Information in Engineering Conference ◽

10.1115/detc2006-99135 ◽

2006 ◽

Author(s):

Eliab Z. Opiyo ◽

Imre Horva´th

Keyword(s):

User Interfaces ◽

Computer Aided Design ◽

3D Imaging ◽

Three Dimensional ◽

3D Display ◽

Desktop Computer ◽

Natural Interaction ◽

Input Devices ◽

3D Space ◽

3D Displays

Standard two-dimensional (2D) computer displays are traditionally used in engineering design to display the three-dimensional (3D) images generated by computer-aided design and engineering (CAD/CAE) systems. These displays serve primarily as passive visualization tools. The interaction with the displayed images on these devices is only possible through archaic 2D peripheral input devices such as keyboards and mice; via the Windows, Icons, Menus and Pointing (WIMP) style graphical user interfaces. It is widely acknowledged in the design community that such visualization and interaction methods do not match the way the designers think and work. Overall, the emerging volumetric 3D displays are seen as the obvious replacement of flat displays in future. This paper explores the possibility of stepping beyond the present 2D desktop computer monitors, and investigate the practicalities of using the emerging volumetric 3D displays, coupled with non encumbering natural interaction means such as gestures, hand motions and haptics for designing in 3D space. We first explore the need for spatial visualization and interaction in design, and outline how the volumetric 3D imaging devices could be used in design. We then review the existing volumetric 3D display configurations, and investigate how they would assist designing in 3D space. Next, we present the study we conducted to seek views of the designers on what kind of volumetric 3D display configuration would more likely match their needs. We finally highlight what would be the consequences and benefits of using volumetric 3D displays instead of the canonical flat screen displays and 2D input devices in design. It has been established that the designers who participated as subjects in the above-mentioned preliminary field study feel that dome-shaped and aerial volumetric 3D imaging devices, which allow for both visualization and interaction with virtual objects, are the imaging options that would not only better suit their visualization and interaction needs, but would also satisfy most of the usability requirements. However, apart from dealing with the remaining basic technological gaps, the challenge is also on how to combine the prevailing proven CAD/CAE technologies and the emerging interaction technologies with the emerging volumetric 3D imaging technologies. As a result of turning to volumetric 3D imaging devices, there is also the challenge of putting in place a formal methodology for designing in 3D space by using these devices.

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Determining androgen inactivation status in prostate cancer by [18F] DHT PET.

Journal of Clinical Oncology ◽

10.1200/jco.2019.37.7_suppl.191 ◽

2019 ◽

Vol 37 (7_suppl) ◽

pp. 191-191

Author(s):

Ziqi Zhu ◽

Yoon-Mi Chung ◽

Olga Sergeeva ◽

Vladimir Kepe ◽

Michael Berk ◽

...

Keyword(s):

Prostate Cancer ◽

Retention Rate ◽

Xenograft Model ◽

Treatment Resistance ◽

Fold Increase ◽

Standard Uptake Value ◽

Retention Rates ◽

Castration Resistant Prostate Cancer ◽

Pet Ct

191 Background: Castration-resistant prostate cancer occurs in part due to increased tumor tissue testosterone (T) and dihydrotestosterone (DHT) that sustain tumor growth. T and DHT are converted to inactive T- and DHT-glucuronide (T/DHT-G) by uridine 5'-diphospho-glucuronosyltransferase family genes (UGT2B15 and UGT2B17) in glucuronidation-competent cells and excreted, but not in glucuronidation-deficient cells. Thus, low glucuronidation activity enables prostate tumors to preserve androgens, which increases hormone treatment resistance and may be detectable by functional imaging. Methods: We knocked out (KO) UGT2B15 and 17 in LNCaP cells and tested the DHT retention rate in the cell lines by pulse-chase using [3H]DHT as a probe. Free and DHT-G retention rates were separately determined after 5 to 60 min. To increase the signal difference between control and KO cells, we screened several ATP-binding cassette transporter inhibitors to block DHT-G excretion. We performed [18F]DHT PET/CT in castrated mice having a control and a KO xenograft on contralateral flanks ( n = 3). The ratio of the standard uptake value (SUV) in control to KO xenografts in each mouse was calculated. To increase the ratio between control and KO tumors, 50µg cyclosporin A (CSA) was injected 30 min before injecting [18F]DHT. Results: After 5 minutes of chase, control cells retained twice the DHT of KO cells. In control cells, 50%-70% DHT was glucuronidated. Almost no DHT-G was detected in KO cells, and free DHT was similar to control. Of the inhibitors, only CSA increased DHT-G (but not free DHT) in control cells, resulting in a 3-4-fold increase in overall signal. In vivo PET/CT showed control xenografts had higher peak SUV but also a higher elution rate. CSA increased the SUV ratio by 1.5-2. Conclusions: We developed a PET/CT modality to detect androgen inactivation in a prostate cancer xenograft model. Androgen-glucuronidation-proficient tumors give off a stronger signal that is increased by ATP transporter inhibition. Our method can provide a noninvasive means of determining androgen metabolism status and therefore could possibly predict effectiveness of potential therapies in a subgroup of tumors predisposed to androgen deprivation resistance.

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