A proteomic investigation of protein adsorption to cerebral microdialysis membranes in surgically treated intracerebral hemorrhage patients

Abstract Background: Cerebral microdialysis (CMD) is a minimally invasive technique for sampling the interstitial fluid in human brain tissue. CMD allows monitoring the metabolic state of tissue, as well as sampling macromolecules such as proteins and peptides. Recovery of proteins or peptides can be hampered by their adsorption to the CMD membrane as has been previously shown in-vitro, however, protein adsorption to CMD membranes has not been characterized following implantation in human brain tissue. Methods: In this paper, we describe the pattern of proteins adsorbed to CMD membranes compared to that of the microdialysate and of cerebrospinal fluid (CSF). We retrieved CMD membranes from three surgically treated intracerebral hemorrhage (ICH) patients, and analyzed protein adsorption to the membranes using two-dimensional gel electrophoresis (2-DE) in combination with nano-liquid mass spectrometry. We compared the proteome profile of three compartments; the CMD membrane, the microdialysate and ventricular CSF collected at time of CMD removal. Results: We found unique protein patterns for each of the three compartments, particularly in the molecular weight range of 10–35 kDa. Conclusion: This study highlights the importance of analyzing the membranes in addition to the microdialysate when using CMD to sample proteins for biomarker investigation.

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A proteomic investigation of protein adsorption to cerebral microdialysis membranes in surgically treated intracerebral hemorrhage patients

10.21203/rs.3.rs-19619/v1 ◽

2020 ◽

Author(s):

Lovisa Tobieson ◽

Zita Czifra ◽

Karin Wåhlén ◽

Niklas Marklund ◽

Bijar Ghafouri

Keyword(s):

Intracerebral Hemorrhage ◽

Human Brain ◽

Protein Adsorption ◽

Brain Tissue ◽

Cerebral Microdialysis ◽

Invasive Technique ◽

Human Brain Tissue ◽

Protein Patterns ◽

Molecular Weight Range ◽

Two Dimensional Gel Electrophoresis

Abstract Background Cerebral microdialysis (CMD) is a minimally invasive technique for sampling the interstitial fluid in human brain tissue. CMD allows monitoring the metabolic state of tissue, as well as sampling macromolecules such as proteins and peptides. Recovery of proteins or peptides can be hampered by their adsorption to the CMD membrane as has been previously shown in-vitro, however, protein adsorption to CMD membranes has not been characterized following implantation in human brain tissue. Methods In this paper, we describe the pattern of proteins adsorbed to CMD membranes compared to that of the microdialysate and of cerebrospinal fluid (CSF). We retrieved CMD membranes from three surgically treated intracerebral hemorrhage (ICH) patients, and analyzed protein adsorption to the membranes using two-dimensional gel electrophoresis (2-DE) in combination with nano-liquid mass spectrometry. We compared the proteome profile of three compartments; the CMD membrane, the microdialysate and ventricular CSF collected at time of CMD removal. Results We found unique protein patterns for each of the three compartments, particularly in the molecular weight range of 10-35 kDa. Conclusion This study highlights the importance of analyzing the membranes in addition to the microdialysate when using CMD to sample proteins for biomarker investigation.

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Proteomic investigation of protein adsorption to cerebral microdialysis membranes in surgically treated intracerebral hemorrhage patients - a pilot study

10.21203/rs.3.rs-25981/v2 ◽

2020 ◽

Author(s):

Lovisa Tobieson ◽

Zita Czifra ◽

Karin Wåhlén ◽

Niklas Marklund ◽

Bijar Ghafouri

Keyword(s):

Intracerebral Hemorrhage ◽

Human Brain ◽

Protein Adsorption ◽

Brain Tissue ◽

Cerebral Microdialysis ◽

Invasive Technique ◽

Human Brain Tissue ◽

Protein Patterns ◽

Molecular Weight Range ◽

Two Dimensional Gel Electrophoresis

Abstract Background: Cerebral microdialysis (CMD) is a minimally invasive technique for sampling the interstitial fluid in human brain tissue. CMD allows monitoring the metabolic state of tissue, as well as sampling macromolecules such as proteins and peptides. Recovery of proteins or peptides can be hampered by their adsorption to the CMD membrane as has been previously shown in-vitro, however, protein adsorption to CMD membranes has not been characterized following implantation in human brain tissue. Methods: In this paper, we describe the pattern of proteins adsorbed to CMD membranes compared to that of the microdialysate and of cerebrospinal fluid (CSF). We retrieved CMD membranes from three surgically treated intracerebral hemorrhage (ICH) patients, and analyzed protein adsorption to the membranes using two-dimensional gel electrophoresis (2-DE) in combination with nano-liquid mass spectrometry. We compared the proteome profile of three compartments; the CMD membrane, the microdialysate and ventricular CSF collected at time of CMD removal. Results: We found unique protein patterns in the molecular weight range of 10-35 kDa for each of the three compartments. Conclusion: This study highlights the importance of analyzing the membranes in addition to the microdialysate when using CMD to sample proteins for biomarker investigation.

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Optimization of the first dimension for separation by two-dimensional gel electrophoresis of basic proteins from human brain tissue

PROTEOMICS ◽

10.1002/pmic.200300624 ◽

2004 ◽

Vol 4 (1) ◽

pp. 27-30 ◽

Cited By ~ 38

Author(s):

Kyla Pennington ◽

Emma McGregor ◽

Clare L. Beasley ◽

Ian Everall ◽

David Cotter ◽

...

Keyword(s):

Human Brain ◽

Brain Tissue ◽

Gel Electrophoresis ◽

Two Dimensional ◽

Human Brain Tissue ◽

Two Dimensional Gel Electrophoresis ◽

Basic Proteins

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Optimization of the first dimension for separation by two-dimensional gel electrophoresis of basic proteins from human brain tissue (Proteomics 2004, vol. 4, issue 1, pp. 27-30)

PROTEOMICS ◽

10.1002/pmic.200490026 ◽

2004 ◽

Vol 4 (5) ◽

pp. 1519-1519

Author(s):

Kyla Pennington ◽

Emma McGregor ◽

Clare L. Beasley ◽

Ian Everall ◽

David Cotter ◽

...

Keyword(s):

Human Brain ◽

Brain Tissue ◽

Gel Electrophoresis ◽

Two Dimensional ◽

Human Brain Tissue ◽

Two Dimensional Gel Electrophoresis ◽

Basic Proteins ◽

Tissue Proteomics

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RBIO-06. IMPLEMENTATION OF HUMAN INDUCED PLURIPOTENT STEM CELL DERIVED CEREBRAL ORGANOIDS TO MODEL NORMAL TISSUE RADIORESPONSE

Neuro-Oncology ◽

10.1093/neuonc/noaa215.806 ◽

2020 ◽

Vol 22 (Supplement_2) ◽

pp. ii193-ii193

Author(s):

Lawrence Bronk ◽

Sanjay Singh ◽

Riya Thomas ◽

Luke Parkitny ◽

Mirjana Maletic-Savatic ◽

...

Keyword(s):

Stem Cell ◽

Human Brain ◽

Brain Tissue ◽

Human Tissue ◽

Model Systems ◽

Human Brain Tissue ◽

Post Irradiation ◽

Mature Neurons ◽

Induced Pluripotent ◽

Effects Of Radiation

Abstract Treatment-related sequelae following cranial irradiation have life changing impacts for patients and their caregivers. Characterization of the basic response of human brain tissue to irradiation has been difficult due to a lack of preclinical models. The direct study of human brain tissue in vitro is becoming possible due to advances in stem cell biology, neuroscience, and tissue engineering with the development of organoids as novel model systems which enable experimentation with human tissue models. We sought to establish a cerebral organoid (CO) model to study the radioresponse of normal human brain tissue. COs were grown using human induced pluripotent stem cells and a modified Lancaster protocol. Compositional analysis during development of the COs showed expected populations of neurons and glia. We confirmed a population of microglia-like cells within the model positive for the makers Iba1 and CD68. After 2-months of maturation, COs were irradiated to 0, 10, and 20 Gy using a Shepard Mark-II Cs-137 irradiator and returned to culture. Subsets of COs were prepared for immunostaining at 30- and 70-days post-irradiation. To examine the effect of irradiation on the neural stem cell (NSC) population, sections were stained for SOX2 and Ki-67 expression denoting NSCs and proliferation respectively. Slides were imaged and scored using the CellProfiler software package. The percentage of proliferating NSCs 30-days post-irradiation was found to be significantly reduced for irradiated COs (5.7% (P=0.007) and 3.4% (P=0.001) for 10 and 20 Gy respectively) compared to control (12.7%). The reduction in the proliferating NSC population subsequently translated to a reduced population of NeuN-labeled mature neurons 70 days post-irradiation. The loss of proliferating NSCs and subsequent reduction in mature neurons demonstrates the long-term effects of radiation. Our initial results indicate COs will be a valuable model to study the effects of radiation therapy on normal and diseased human tissue.

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