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Highly dimensional single cell analysis of peripheral inflammation in ALS with mass cytometry

Messing, Melina (2017) MOBT01 20162
Degree Projects in Molecular Biology
Abstract
Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease with neither a known cause nor an available cure. With disease progression, toxic protein aggregates form in the brain, leading to the loss of the upper and lower motor neurons. While circulating bone marrow-derived inflammatory cells have not been shown to cross the blood brain barrier, changes in their number and composition may still be seen during disease progression. The sporadic nature and variable phenotype as well as lack of a definitive biomarker prevent a fast and definite diagnosis and underline the necessity for a deep understanding of the peripheral inflammatory changes.

In order to broadly investigate peripheral inflammation, mononuclear cell... (More)
Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease with neither a known cause nor an available cure. With disease progression, toxic protein aggregates form in the brain, leading to the loss of the upper and lower motor neurons. While circulating bone marrow-derived inflammatory cells have not been shown to cross the blood brain barrier, changes in their number and composition may still be seen during disease progression. The sporadic nature and variable phenotype as well as lack of a definitive biomarker prevent a fast and definite diagnosis and underline the necessity for a deep understanding of the peripheral inflammatory changes.

In order to broadly investigate peripheral inflammation, mononuclear cell fractions from ALS patients and controls were obtained and phenotypically characterized using the highly dimensional single cell analysis technology mass cytometry and a panel of 35 metal-tagged antibodies specific for inflammatory surface markers. Peripheral blood phenotypes were then compared to ALS-FRS data to correlate clinical symptoms with changes in cell subsets. Data analysis was performed with a combination of FlowJo, Cytobank (CITRUS) and Cytofkit (Phenograph).

At the time of writing, 19 patients and 19 control samples were processed for mass cytometry and following stage-based patient grouping several populations preliminarily could be associated with different stages of ALS.
Among these, a myeloid population was extensively characterized by CD31, CD14, CD11b, CD11c, CD4, TLR2, and CCR2 and was specifically increased for stage 3 (lower limb onset) patients. Additionally, activated T helper cells (CD4+, CD27+, CD28+) showed an increase in numbers at late stage ALS. Changes in marker expression showed variability both between patients and in comparison to day-matched controls.

Despite sample variability and limited sample number, this study has laid the groundwork for a detailed characterization of peripheral inflammation in ALS and demonstrated the power of highly dimensional single cell analysis by mass cytometry.
Building on these findings, the data holds potential to identify ALS specific biomarkers to enhance our understanding of ALS and to facilitate diagnosis, prognosis and to predict effectiveness in clinical trials as well as identify potential targets for therapeutic treatments. (Less)
Popular Abstract
Peripheral Inflammation in ALS

Highly dimensional single cell analysis was performed with mass cytometry in order to broadly characterize peripheral inflammation in ALS patients and to find disease specific as well as disease stage specific cellular signatures.

Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease with neither a known cause nor an available cure. With disease progression, toxic protein aggregates form in the brain, leading to the loss of the upper and lower motor neurons. While circulating bone marrow-derived inflammatory cells have not been shown to cross the blood brain barrier, changes in their number and compositions have been observed both in rodent models and ALS patients.
The... (More)
Peripheral Inflammation in ALS

Highly dimensional single cell analysis was performed with mass cytometry in order to broadly characterize peripheral inflammation in ALS patients and to find disease specific as well as disease stage specific cellular signatures.

Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease with neither a known cause nor an available cure. With disease progression, toxic protein aggregates form in the brain, leading to the loss of the upper and lower motor neurons. While circulating bone marrow-derived inflammatory cells have not been shown to cross the blood brain barrier, changes in their number and compositions have been observed both in rodent models and ALS patients.
The sporadic nature and variable phenotype as well as lack of a definitive biomarker prevent a fast and definite diagnosis and underline the necessity for a deep understanding of the peripheral inflammatory changes.

In order to broadly investigate peripheral inflammation, mononuclear cell fractions from ALS patients and controls were obtained and phenotypically characterized using mass cytometry and a panel of 35 metal-tagged antibodies specific for inflammatory surface markers. Peripheral blood phenotypes were then compared to ALS-FRS data to correlate clinical symptoms with changes in cell subsets. Data analysis was performed with a combination of FlowJo, Cytobank (CITRUS) and Cytofkit (Phenograph).

At the time of writing, 19 patients and 19 control samples were processed for mass cytometry and following stage-based patient grouping, several populations preliminarily could be associated with different stages of disease. Among these, a myeloid population was extensively characterized by CD31, CD14, CD11b, CD11c, CD4, TLR2, and CCR2 and was specifically increased for stage 3 (lower limb onset) patients. Additionally, activated T helper cells (CD4+, CD27+, CD28+) showed an increase in numbers at late stage ALS. Changes in marker expression showed variability both between patients and in comparison to day-matched controls.

Despite sample variability and limited sample number, this study has laid the groundwork for a detailed characterization of peripheral inflammation in ALS and demonstrated the power of highly dimensional single cell analysis by mass cytometry.
Building on these findings, the data holds potential to identify ALS specific biomarkers to enhance our understanding of ALS and to facilitate diagnosis, prognosis and to predict effectiveness in clinical trials as well as identify potential targets for therapeutic treatments.

Master’s Degree Project in Molecular Biology 60 credits 2017
Department of Biology, Lund University

Advisor: Dr. Fabio Rossi (UBC), Dr. Marita Cohn (LU)
Department of Medical Genetics, University of British Columbia,
Department of Biology, Lund University (Less)
Please use this url to cite or link to this publication:
author
Messing, Melina
supervisor
organization
course
MOBT01 20162
year
type
H2 - Master's Degree (Two Years)
subject
language
English
id
8925232
date added to LUP
2017-09-11 09:48:08
date last changed
2017-09-11 09:49:02
@misc{8925232,
  abstract     = {Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease with neither a known cause nor an available cure. With disease progression, toxic protein aggregates form in the brain, leading to the loss of the upper and lower motor neurons. While circulating bone marrow-derived inflammatory cells have not been shown to cross the blood brain barrier, changes in their number and composition may still be seen during disease progression. The sporadic nature and variable phenotype as well as lack of a definitive biomarker prevent a fast and definite diagnosis and underline the necessity for a deep understanding of the peripheral inflammatory changes. 

In order to broadly investigate peripheral inflammation, mononuclear cell fractions from ALS patients and controls were obtained and phenotypically characterized using the highly dimensional single cell analysis technology mass cytometry and a panel of 35 metal-tagged antibodies specific for inflammatory surface markers. Peripheral blood phenotypes were then compared to ALS-FRS data to correlate clinical symptoms with changes in cell subsets. Data analysis was performed with a combination of FlowJo, Cytobank (CITRUS) and Cytofkit (Phenograph).

At the time of writing, 19 patients and 19 control samples were processed for mass cytometry and following stage-based patient grouping several populations preliminarily could be associated with different stages of ALS.
Among these, a myeloid population was extensively characterized by CD31, CD14, CD11b, CD11c, CD4, TLR2, and CCR2 and was specifically increased for stage 3 (lower limb onset) patients. Additionally, activated T helper cells (CD4+, CD27+, CD28+) showed an increase in numbers at late stage ALS. Changes in marker expression showed variability both between patients and in comparison to day-matched controls.

Despite sample variability and limited sample number, this study has laid the groundwork for a detailed characterization of peripheral inflammation in ALS and demonstrated the power of highly dimensional single cell analysis by mass cytometry.
Building on these findings, the data holds potential to identify ALS specific biomarkers to enhance our understanding of ALS and to facilitate diagnosis, prognosis and to predict effectiveness in clinical trials as well as identify potential targets for therapeutic treatments.},
  author       = {Messing, Melina},
  language     = {eng},
  note         = {Student Paper},
  title        = {Highly dimensional single cell analysis of peripheral inflammation in ALS with mass cytometry},
  year         = {2017},
}