Download PDF
Phlebologie 2020; 49(04): 242-248
DOI: 10.1055/a-1191-8380
Originalarbeit

Light sheet microscopy-based 3-dimensional histopathology of the lymphatic vasculature in Emberger syndrome

Article in several languages: English | deutsch
René Hägerling
  Institut für Medizinische Genetik und Humangenetik, Charité-Universitätsmedizin Berlin
› Author Affiliations
› Further Information
   Permissions and Reprints

Abstract

Introduction Lymphovascular diseases represent a heterogenous group of inherited and sporadic disorders and refer to a range of possible underlying pathologies and pathogenesis.

Emberger Syndrome, an inherited form of lymphedema, is characterized by bilateral lower limb lymphedema, however, upper limbs do not show any signs of swelling.

To identify disease-associated histopathological alterations in patients with Emberger Syndrome and to elucidate potential histological differences between the lymphatic vasculature of upper and lower limbs, a detailed knowledge on the 3-dimensional tissue and vessel architecture is essential. However, the current gold standard in 2-dimensional histology provides only very limited spatial information.

Material and methods To elucidate the underlying vascular pathology in Emberger Syndrome on the cellular level, we applied the 3-dimensional visualization and analysis approach VIPAR (volume information-based histopathological analysis by 3D reconstruction and data extraction) to entire wholemount immunofluorescence-stained human tissue samples. VIPAR is a light sheet microscopy-based imaging technique, which allows 3-dimensional reconstruction of entire tissue biopsies followed by automated and semi-automated analysis of vascular parameters in 3-dimensional space.

Results Using VIPAR we could show that in Emberger Syndrome the dermal lymphatic vasculature is intact and non-disrupted.

However, lower limbs showed an hypoplastic lymphatic vasculature with absence of lymphatic valves in pre-collecting and collecting vessels. In contrast to the lower limbs, the lymphatic vasculature of the upper limbs showed no morphological alterations of lymphatic vessels and lymphatic valves compared to healthy controls.

Discussion Based on the 3-dimensional histopathological analysis we were able to perform a detailed phenotyping of lymphatic vessels in the upper and lower limb in Emberger Syndrome and to identify the underlying vascular pathology. In addition, we could show vascular alteration between the upper and lower limbs indicating a vascular heterogeneity of dermal lymph vessels causing the lower limb lymphedema.

Key words

light sheet microscopy - histopathology - lymphovascular diseases - lymphoedema

Schlüsselwörter

Lichtblattmikroskopie - Histopathologie - Lymphovaskuläre Erkrankungen - Lymphödem


Publication History

Article published online:
21 August 2020

© Georg Thieme Verlag KG
Stuttgart · New York

 

  • References

  • 1 Mortimer PS, Rockson SG. New developments in clinical aspects of lymphatic disease. J Clin Invest 2014; 124 (03) 915-921
    CrossrefPubMedGoogle Scholar
  • 2 Schulte-Merker S, Sabine A, Petrova TV. Lymphatic vascular morphogenesis in development, physiology, and disease. J Cell Biol 2011; 193 (04) 607-618
    CrossrefPubMedGoogle Scholar
  • 3 Ostergaard P, Simpson MA, Connell FC. Mutations in GATA2 cause primary lymphedema associated with a predisposition to acute myeloid leukemia (Emberger syndrome). Nat Genet 2011; 42 (10) 929-931
    CrossrefPubMedGoogle Scholar
  • 4 Kazenwadel J, Secker GA, Liu YJ. Loss-of-function germline GATA2 mutations in patients with MDS/AML or MonoMAC syndrome and primary lymphedema reveal a key role for GATA2 in the lymphatic vasculature. Blood 2012; 119 (05) 1283-1291
    CrossrefPubMedGoogle Scholar
  • 5 Donadieu J, Lamant M, Fieschi C. Natural history of GATA2 deficiency in a survey of 79 French and Belgian patients. Haematologica 2018; 103 (08) 1278-1287
    CrossrefPubMedGoogle Scholar
  • 6 Brambila-Tapia AJL, García-Ortiz JE, Brouillard P. GATA2 Null Mutation Associated With Incomplete Penetrance in a Family With Emberger Syndrome. Hematology 2017; 22 (08) 467-471
    PubMedGoogle Scholar
  • 7 Kazenwadel J, Betterma KL, Chong CE. GATA2 is required for lymphatic vessel valve development and maintenance. J Clin Invest 2015; 125 (08) 2979-2994
    CrossrefPubMedGoogle Scholar
  • 8 Mahamud MR, Geng X, Ho YC. GATA2 controls lymphatic endothelial cell junctional integrity and lymphovenous valve morphogenesis through miR-126. Development 2019; 146 (21) dev184218
    CrossrefPubMedGoogle Scholar
  • 9 Cha B, Geng X, Mahamud MR. Complementary Wnt Sources Regulate Lymphatic Vascular Development via PROX1-Dependent Wnt/β-Catenin Signaling. Cell Rep 2018; 25 (03) 571-584.e5
    CrossrefPubMedGoogle Scholar
  • 10 Greene AK, Grant FD, Slavin SA. Lower-extremity lymphedema and elevated body-mass index. N Engl J Med 2012; 366 (22) 2136-2137
    CrossrefPubMedGoogle Scholar
  • 11 Connell FC, Gordon K, Brice G. The classification and diagnostic algorithm for primary lymphatic dysplasia: an update from 2010 to include molecular findings. Clin Genet 2013; 84 (04) 303-314
    CrossrefPubMedGoogle Scholar
  • 12 Hägerling R, Drees D, Scherzinger A. VIPAR, a quantitative approach to 3D histopathology applied to lymphatic malformations. JCI Insight 2017; 2 (16) e93424
    CrossrefPubMedGoogle Scholar
  • 13 Hägerling R, Pollmann C, Andreas M. A novel multistep mechanism for initial lymphangiogenesis in mouse embryos based on ultramicroscopy. EMBO J 2013; 32 (05) 629-644
    CrossrefPubMedGoogle Scholar
  • 14 Pollmann C, Hägerling R, Kiefer F. Visualization of lymphatic vessel development, growth, and function. Adv Anat Embryol Cell Biol 2014; 214: 167-186
    CrossrefPubMedGoogle Scholar
  • 15 Wigle JT, Oliver G. Prox1 function is required for the development of the murine lymphatic system. Cell 1999; 98 (06) 769-778
    CrossrefPubMedGoogle Scholar