endometriosi alle porte del nuovo decennio auditorium museo … · 2020-04-29 · roberta bulla,...
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Roberta Bulla, PhD
Laboratory of Immunology
Deparment of Life Sciences
University of Trieste
ITALY
Endometriosi alle porte del nuovo decennio
13/12/2019Auditorium Museo Revoltella
– Via Diaz,27 – Trieste
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Introduction
pelvic paindysmenorrheadyspareunia dysuria
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Introduction
� Characterized by the presence of functional endometrial tissue outside theuterine cavity.
� The hormonal cycle iduces the bleeding of this ectopic tissue leading to achronic inflammatory condition.
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All the proposed hypothesies for the cell origincan be categorized into two main theory� In situ theory� Transplantation theory
Introduction
Retrograde Menstruation Coelomic Metaplasia
Müllerian Theory or Defective Embryogenesis
Stem Cell Theory
Laganà IJMS 2019 doi:10.3399/ijms20225615
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Stroma and glands of endometrial-like tissue of endometriosis originate in-situ from the local tiss ue by
metaplasia or by embryological origin
Introduction
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Stroma and glands of endometrial-like tissue of endometriosis originate from eutopic endometrium.
Endometriosis is proposed as a benign metastasis of eutopic endometrium which is displaced from the uterine cavity to another location inside the body
through different routes: hematogenous, lymphatic a nd iatrogenic (mechanical) spread of endometrial or
endometriotic cells
Introduction
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Retrograde menstruation and
immune disfunction
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Endometriotic lesion immune
microenvironment
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Mast cells mediate inflammation
and fibrosis in endometriosis
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Mast cells mediate inflammation
and fibrosis in endometriosis
EMJ Repro Health. 2017;3[1]:76-83
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Complement System Pathway is
alterated in EM
introductionSuryewanshi S. et al., Clinical Cancer Research 2014
Gene expression analysis revealed the complement pathway as mostprominently involved in endometriosis
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Complement cascade ActivationAlternative way
C3H2O to several activating surfaces
Lectinin wayMBL, fycolins or
collectin 11 bind to carbohydrates
Classical wayC1q binds to
immunocomplexes
CYTOLYSIS
MAC sTCC
INFLAMMATION
Brai, 1991
INFLAMMATION
OPSONIZATION
C3b, C3bi, C4b
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Several groups* demonstrated that the glandularepithelial and stromal cells found in endometrioticimplants produce and secrete the C component C3.
The aim of this work was to confirm the presence ofC3 in the ectopic tissue in comparison to the eutopicone, and to investigate the direct role of C3 in thepathogenesis of EM.
*Weed and Arquembourg 1980; Bartosik D. et al., 1987; Isaacson K.B. et al.,1989; Bischof P. et al., 1994; Ruiz LA et al., 2011; Signorile P.G. et al., 2014,Suryawanshi S. et al., 2014; Rekker et al., 2017
AIM
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AEC
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-RESULTS-
n = 4
n = 3
endometriotic cyst tissue presents higher level of C3 (but not C4 or C5) mRNA compared to normal uterus.
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-RESULTS-
n = 4
n = 3
Endometriotic cells isolated from cysts and cultured in vitro, were able to synthesized C3.
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-Results-
Endometrial cells, when
stimulated with pro-
inflammatory stimuli (in
particular with TNF-α) start
to express C3
(but not C4 or C5)
AN3CA (human endometrial cell line)
MW120
75
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The engraftment and the
dimensions of
endometriotic lesions is
reduced in C3 deficient mice
compared to WT mice
-RESULTS-
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-RESULTS-
CTRL - Untreated WT C3 KO
WT
Mou
se P
erito
enal
was
hing
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-Results-
The analysis of the peritoneal liquids, collected from endometriotic mice, reveledan increase of tryptase and β-hexosaminidase, enzymes present in the mast-cellgranules, in WT animals compared to C3-/- mice.
*
*
*
Tryptase Beta-hexosaminidase
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Results
toluidine blue
CTRL from explorative laparoscopy
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Peritoneal liquid from endometriotic
patients stimulated mast cells to produce
proinflammatory cytokines
C3 mRNA expression by AN3CA
Restin
g αααα
TNF-
EMPL
HMC-1+E
MPL
0
1
2
3
4
mR
NA
C3/
mR
NA
18S
Endometrial cells
mast cells
Endometrioticperitoneal liquid
C3
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C3
Mast-Cells
Endometrial tissue
-Conclusions-
C3a
Cytokines
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Endometriosis treatment
Sodiumcromoglycate
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Endometriosis treatment
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C3a antagonists
Endometriosis treatment
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Endometriosis treatment
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Department of Life Sciences
C. AgostinisS. ZorzetF. BossiP. ZacchiA. BalduitA. MangognaV. BorelliF. Tedesco
UNIVERSITY OF TRIESTE
G. RicciG. ZitoF. RomanoO. Radillo
IRCCS BURLO GAROFOLO
M. Botto
Department of Obstetrics and Gynaecology
IMPERIAL COLLEGELondon, UK
UNIVERSITY OF PALERMOC. TripodoB. BelmonteA. Gulino
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Thank you for your attention
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� The complement component C3 is locally synthetized by ectopicendometrial tissue.
� Normal endometrial cells under pro-inflammatory stimuli areable to produce C3
� C3 deficient mice present less endometriotic lesions in asyngeneic EM mouse model
-CONCLUSIONS-
C3 is involved in the pathogenesis of EM
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10x 10x
20x 20x