TY - JOUR
T1 - Quantifying lymphocyte vacuolization serves as a measure of CLN3 disease severity
AU - Kuper, Willemijn F.E.
AU - Oostendorp, Marlies
AU - van den Broek, Brigitte T.A.
AU - van Veghel, Karin
AU - Nonkes, Lourens J.P.
AU - Nieuwenhuis, Edward E.S.
AU - Fuchs, Sabine A.
AU - Veenendaal, Tineke
AU - Klumperman, Judith
AU - Huisman, Albert
AU - Nierkens, Stefan
AU - van Hasselt, Peter M.
N1 - Publisher Copyright:
© 2020 The Authors. Journal of Inherited Metabolic Disease published by John Wiley & Sons Ltd on behalf of SSIEM.
PY - 2020/1/1
Y1 - 2020/1/1
N2 - Background: The CLN3 disease spectrum ranges from a childhood-onset neurodegenerative disorder to a retina-only disease. Given the lack of metabolic disease severity markers, it may be difficult to provide adequate counseling, particularly when novel genetic variants are identified. In this study, we assessed whether lymphocyte vacuolization, a well-known yet poorly explored characteristic of CLN3 disease, could serve as a measure of disease severity. Methods: Peripheral blood obtained from healthy controls and CLN3 disease patients was used to assess lymphocyte vacuolization by (a) calculating the degree of vacuolization using light microscopy and (b) quantifying expression of lysosomal-associated membrane protein 1 (LAMP-1), using flow cytometry in lymphocyte subsets as well as a qualitative analysis using electron microscopy and ImageStream analysis. Results: Quantifying lymphocyte vacuolization allowed to differentiate between CLN3 disease phenotypes (P =.0001). On immunofluorescence, classical CLN3 disease lymphocytes exhibited abundant vacuole-shaped LAMP-1 expression, suggesting the use of LAMP-1 as a proxy for lymphocyte vacuolization. Using flow cytometry in lymphocyte subsets, quantifying intracellular LAMP-1 expression additionally allowed to differentiate between infection and storage and to differentiate between CLN3 phenotypes even more in-depth revealing that intracellular LAMP-1 expression was most pronounced in T-cells of classical-protracted CLN3 disease while it was most pronounced in B-cells of “retina-only” CLN3 disease. Conclusion: Lymphocyte vacuolization serves as a proxy for CLN3 disease severity. Quantifying vacuolization may help interpretation of novel genetic variants and provide an individualized readout for upcoming therapies.
AB - Background: The CLN3 disease spectrum ranges from a childhood-onset neurodegenerative disorder to a retina-only disease. Given the lack of metabolic disease severity markers, it may be difficult to provide adequate counseling, particularly when novel genetic variants are identified. In this study, we assessed whether lymphocyte vacuolization, a well-known yet poorly explored characteristic of CLN3 disease, could serve as a measure of disease severity. Methods: Peripheral blood obtained from healthy controls and CLN3 disease patients was used to assess lymphocyte vacuolization by (a) calculating the degree of vacuolization using light microscopy and (b) quantifying expression of lysosomal-associated membrane protein 1 (LAMP-1), using flow cytometry in lymphocyte subsets as well as a qualitative analysis using electron microscopy and ImageStream analysis. Results: Quantifying lymphocyte vacuolization allowed to differentiate between CLN3 disease phenotypes (P =.0001). On immunofluorescence, classical CLN3 disease lymphocytes exhibited abundant vacuole-shaped LAMP-1 expression, suggesting the use of LAMP-1 as a proxy for lymphocyte vacuolization. Using flow cytometry in lymphocyte subsets, quantifying intracellular LAMP-1 expression additionally allowed to differentiate between infection and storage and to differentiate between CLN3 phenotypes even more in-depth revealing that intracellular LAMP-1 expression was most pronounced in T-cells of classical-protracted CLN3 disease while it was most pronounced in B-cells of “retina-only” CLN3 disease. Conclusion: Lymphocyte vacuolization serves as a proxy for CLN3 disease severity. Quantifying vacuolization may help interpretation of novel genetic variants and provide an individualized readout for upcoming therapies.
KW - CLN3 disease
KW - flow cytometry
KW - ImageStream
KW - lymphocyte vacuolization
KW - lysosomal membrane-associated protein-1 (LAMP-1)
KW - neuronal ceroid lipofuscinosis (NCL)
UR - http://www.scopus.com/inward/record.url?scp=85092606783&partnerID=8YFLogxK
U2 - 10.1002/jmd2.12128
DO - 10.1002/jmd2.12128
M3 - Article
AN - SCOPUS:85092606783
SN - 2192-8304
VL - 54
SP - 87
EP - 97
JO - JIMD Reports
JF - JIMD Reports
IS - 1
ER -