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. 2018 May 8;19(1):83.
doi: 10.1186/s12931-018-0797-9.

Immunohistological features related to functional impairment in lymphangioleiomyomatosis

Ellen Caroline Toledo do Nascimento  1 , Bruno Guedes Baldi  2 , Alessandro Wasum Mariani  3 , Raquel Annoni  4 , Ronaldo Adib Kairalla  2 , Suzana Pinheiro Pimenta  2 , Luiz Fernando Ferraz da Silva  4 , Carlos Roberto Ribeiro Carvalho  2 , Marisa Dolhnikoff  4
Affiliations

Immunohistological features related to functional impairment in lymphangioleiomyomatosis

Ellen Caroline Toledo do Nascimento et al. Respir Res. .

Abstract

Background: Lymphangioleiomyomatosis (LAM) is a low-grade neoplasm characterized by the pulmonary infiltration of smooth muscle-like cells (LAM cells) and cystic destruction. Patients usually present with airway obstruction in pulmonary function tests (PFTs). Previous studies have shown correlations among histological parameters, lung function abnormalities and prognosis in LAM. We investigated the lung tissue expression of proteins related to the mTOR pathway, angiogenesis and enzymatic activity and its correlation with functional parameters in LAM patients.

Methods: We analyzed morphological and functional parameters of thirty-three patients. Two groups of disease severity were identified according to FEV1 values. Lung tissue from open biopsies or lung transplants was immunostained for SMA, HMB-45, mTOR, VEGF-D, MMP-9 and D2-40. Density of cysts, density of nodules and protein expression were measured by image analysis and correlated with PFT parameters.

Results: There was no difference in the expression of D2-40 between the more severe and the less severe groups. All other immunohistological parameters showed significantly higher values in the more severe group (p ≤ 0.002). The expression of VEGF-D, MMP-9 and mTOR in LAM cells was associated with the density of both cysts and nodules. The density of cysts and nodules as well as the expression of MMP-9 and VEGF-D were associated with the impairment of PFT parameters.

Conclusions: Severe LAM represents an active phase of the disease with high expression of VEGF-D, mTOR, and MMP-9, as well as LAM cell infiltration. Our findings suggest that the tissue expression levels of VEGF-D and MMP-9 are important parameters associated with the loss of pulmonary function and could be considered as potential severity markers in open lung biopsies of LAM patients.

Keywords: Immunohistochemistry; Lymphangioleiomyomatosis; Metalloproteinase; Pulmonary function tests; VEGF-D; mTOR.

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Conflict of interest statement

Ethics approval and consent to participate

The study was approved by the review board for the human ethics committee of Sao Paulo University (CAPPesq-FMUSP n° 0623/09). The study is retrospective and used medical records and archived material from the Department of Pathology of Sao Paulo University Medical School.

Competing interests

The authors declare that they have no competing interests.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
Representative LAM lesion stained at H&E (a). Panel b shows a higher magnification of the typical LAM cells nodule. Cy = cystic component; arrow indicates the cellular component (LAM cells nodule) localized at the cyst wall. Magnifications: A = 70X, B = 200X
Fig. 2
Fig. 2
Representative photomicrographs of LAM lesions extracted for image analysis. Panel a shows an SMA-stained lung biopsy with multiple LAM lesions. Panels b to g show examples of extracted lesions from different immunohistochemical stains. b - SMA, c - HMB45, d - MMP9, e - VEGFD, f - mTOR, g - D2-40. Magnifications: A = 10X, B to G = 40X
Fig. 3
Fig. 3
Representative image analysis. Panel a shows an SMA-stained extracted LAM lesion. Panel b shows the positively stained area (red) at image analysis. Panel c shows the internal perimeter (arrow pointing to the blue perimeter line) of the cystic component of a LAM lesion (H&E staining). Magnification: 40X
Fig. 4
Fig. 4
The graphs show the variability in the number of LAM lesions (a), density of cysts (b), and density of nodules (c) among cases in the two groups
Fig. 5
Fig. 5
Representative histological images of H&E staining and immunohistochemistry for protein expression (stained in brown) in each group. All immunohistological parameters showed significantly higher values in the more severe group. a, c, e, g, i, and k: the less severe group; b, d, f, h, j, and l: the more severe group. A and B: H&E staining showing the distribution of cysts and LAM-cells nodules (arrows) in each group (magnification: 5×). c and d: SMA expression in LAM-cells (magnification: C, 7× and D, 7×). e and f: HMB-45 expression in LAM-cells (magnification: E, 20× and F, 40×). g and h: VEGF-D expression in LAM-cells (magnification: G, 100× and H, 100×). i and j: MMP-9 expression in LAM-cells (magnification: I, 25× and J, 25×). k and l: mTOR expression in LAM-cells (magnification: K, 35× and L, 35×)
Fig. 6
Fig. 6
Kaplan-Meier estimates of freedom from death or lung transplantation for tissue expression of VEGF-D (a), mTOR (b), and MMP-9 (c). For each marker, two subgroups of patients were defined according to the marker median values
See this image and copyright information in PMC

References

    1. Travis WD, Brambilla E, Nicholson AG, et al. The 2015 world health organization classification of lung tumors: impact of genetic, clinical and radiologic advances since the 2004 classification. J Thorac Oncol. 2015;10(9):1243–1260. doi: 10.1097/JTO.0000000000000630. - DOI - PubMed
    1. Travis WD, Brambilla E, Burke AP, Marx A, Nicholson AG. WHO classification of tumours of the lung, pleura, thymus and heart. 4. Lyon, FR: IARC; 2015. - PubMed
    1. McCormack FX, Travis WD, Colby TV, et al. Lymphangioleiomyomatosis: calling it what it is: a low-grade, destructive, metastasizing neoplasm. Am J Respir Crit Care Med. 2012;186(12):1210–1212. doi: 10.1164/rccm.201205-0848OE. - DOI - PMC - PubMed
    1. Hohman DW, Noghrehkar D, Ratnayake S. Lymphangioleiomyomatosis: a review. Eur J Intern Med. 2008;19(5):319–324. doi: 10.1016/j.ejim.2007.10.015. - DOI - PubMed
    1. Baldi BG, Freitas CS, Araujo MS, et al. Clinical course and characterization of lymphangioleiomyomatosis in a brazilian reference centre. Sarcoidosis Vasc Diffuse Lung Dis. 2014;31(2):129–135. - PubMed

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