Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Oct 21;26(11):108262.
doi: 10.1016/j.isci.2023.108262. eCollection 2023 Nov 17.

Th1/17 polarization and potential treatment by an anti-interferon-γ DNA aptamer in Hunner-type interstitial cystitis

Yoshiyuki Akiyama  1   2 , Kaori Harada  3 , Jimpei Miyakawa  1 , Karl J Kreder  2 , Michael A O'Donnell  2 , Maeda Daichi  4 , Hiroto Katoh  5 , Miyuki Hori  3 , Kensuke Owari  3 , Kazunobu Futami  3 , Shumpei Ishikawa  5 , Tetsuo Ushiku  6 , Haruki Kume  1 , Yukio Homma  1   7 , Yi Luo  2
Affiliations

Th1/17 polarization and potential treatment by an anti-interferon-γ DNA aptamer in Hunner-type interstitial cystitis

Yoshiyuki Akiyama et al. iScience. .

Abstract

Hunner-type interstitial cystitis (HIC) is a rare, enigmatic inflammatory disease of the urinary bladder with no curative treatments. In this study, we aimed to characterize the unique cellular and immunological factors specifically involved in HIC by comparing with cystitis induced by Mycobacterium bovis bacillus Calmette-Guérin, which presents similar clinicopathological features to HIC. Here, we show that T helper 1/17 +polarized immune responses accompanied by prominent overexpression of interferon (IFN)-γ, enhanced cGAS-STING cytosolic DNA sensing pathway, and increased plasma cell infiltration are the characteristic inflammatory features in HIC bladder. Further, we developed a mouse anti-IFN-γ DNA aptamer and observed that the intravesical instillation of the aptamer significantly ameliorated bladder inflammation, pelvic pain and voiding dysfunction in a recently developed murine HIC model with little migration into the blood. Our study provides the plausible basis for the clinical translation of the anti-IFN-γ DNA aptamer in the treatment of human HIC.

Keywords: Disease; Immunology.

PubMed Disclaimer

Conflict of interest statement

This study was financially supported by TAGCyx Biotechnologies Inc.

Figures

None
Graphical abstract
Figure 1
Figure 1
Hierarchical clustering analysis of bladder DEGs indicates distinct gene expression patterns between HIC and BCG groups A heatmap generated by hierarchical clustering analysis of DEGs among Hunner lesions, non-Hunner lesion areas, and BCG cystitis. Red: high relative expression; green: low relative expression. See also Table S2.
Figure 2
Figure 2
Bladder quantitative PCR analysis shows increased mRNA levels of pro-Th1/17, cGAS and STING in HIC (A) Quantitative PCR analysis of IL2, IL4, IL6, IL10, IL12A, IL12B, IL17A, IL22, IL23R, HIF1A, IFNA17, IFNB1, IFNG, TNF, TGFB1, TBX21, GATA3, RORC, CTLA4, CGAS, and STING1 in the bladder samples of Hunner lesion areas (HIC-L), non-Hunner lesion areas (HIC-NL), and BCG cystitis. The expression level of each gene is reported relative to that of IPO8. Data shown are the mean ± SD relative gene expressions. (B) Ratios of Th1/Th2 markers (left two graphs) and Th17/Th2 markers (right two graphs) in the bladder samples of Hunner lesion areas (HIC-L), non-Hunner lesion areas (HIC-NL), and BCG cystitis. Data are shown as mean ± SD. ∗p < 0.05 and ∗∗p < 0.01, statistically significant by Steel–Dwass test.
Figure 3
Figure 3
Bladder immunohistochemical quantification demonstrates comparable lymphoplasmacytic and mast cell densities between HIC and BCG cystitis but higher IL-17A-positive cell density and plasma cell ratio in HIC (A) Densities of lymphoplasmacytic (the sum of CD3−, CD20−and CD138-positive cells) and CD4−, CD8−, and mast cell tryptase-positive cells. Representative hematoxylin and eosin staining images of the Hunner lesion areas (HIC-L), non-Hunner lesion areas (HIC-NL), and BCG cystitis are shown along with the lymphoplasmacytic cell density diagram. (scale bar: 250 μm). (B) Densities of IL-17A- and FOXP3-positive cells with representative immunohistochemical images. (scale bar: 200 μm in IL-17A, 100 μm in FOXP3). (C) The ratios of CD4-positivity to CD8-positivity, CD138-positivity to the sum of CD3−, CD20−and CD138-positivity (“Plasma cell ratio”), IL-17A-positivity to FOXP3-positivity, and epithelial area to whole specimen area, with representative immunohistochemical images. (scale bar: 250 μm). Values are expressed as the median (interquartile range). ∗p < 0.05, statistically significant by Steel–Dwass test.
Figure 4
Figure 4
Representative images of serial sections of the bladders from patients with HIC or BCG cystitis evaluated by hematoxylin and eosin staining or immunohistochemical staining with antibodies against CD3, CD4, CD8, CD20, CD138, mast cell tryptase, IL-17A, IFN-γ, cGAS, and STING (A) Hunner lesion of HIC. (B) Non-Hunner lesion area of HIC. (C) BCG cystitis. (a) Hematoxylin and eosin staining. (b–k) Immunohistochemistry: (b) CD3, (c) CD4, (d) CD8, (e) CD20, (f) CD138, (g) mast cell tryptase, (h) IL-17A, (i) IFN-γ, (j) cGAS, (k) STING (scale bar: 200 μm). (l–o) Enlarged images of the areas indicated by the rectangular boxes in h-k, respectively (scale bar: 100 μm).
Figure 5
Figure 5
Bladder immunohistochemical quantification demonstrates increased protein levels of IFN-γ, cGAS and STING in HIC The immunoreactive intensities of IFN-γ, cGAS and STING were evaluated based on the cellular (and stromal in IFN-γ) immunoreactive intensity as follows: negative, mild, moderate, and strong. Specimens with a negative or mild intensity were defined as having low immunoreactivity, and those with a moderate or strong intensity were defined as having high immunoreactivity. (A) Higher immunoreactive intensities of cGAS and STING in both the Hunner lesion area (HIC-L) and non-Hunner lesion area (HIC-NL) of HIC compared to those in BCG cystitis. (B) Higher immunoreactive intensity of IFN-γ in both the Hunner lesion area (a, d) and non-Hunner lesion area (b, e) of HIC, compared to that in BCG cystitis (c, f) with representative immunohistochemical images. (scale bar: 250 μm). (d-f) Enlarged images of the areas indicated by the rectangular boxes in a-c, respectively. (scale bar: 100 μm). ∗p < 0.05 and ∗∗p < 0.01, statistically significant by Fisher’s exact test.
Figure 6
Figure 6
Intravesical instillation of mIFNγ6-1mh significantly reduced bladder inflammation, pelvic nociceptive response, and urinary frequency in the HIC-like URO-OVA cystitis model (A) Experimental outline. (B) Representative bladder histological images. PBS-treated cystitis mice (n = 5) showed dense mononuclear cell infiltration (arrows), increased vascularity (asterisks), mucosal hyperemia (arrowheads), and interstitial edema (horizontal double asterisks). No or mild changes were observed in cystitis-uninduced (n = 5) and mIFNγ6-1mh-treated cystitis mice (n = 7). Magnification: 40× (scale bar: 500 μm), 100× (scale bar: 200 μm) and 400× (scale bar: 50 μm). (C) RT-PCR analysis of bladder Ifng, Tnf, Tac1 and Ngf mRNAs. Top panel: Electrophoresis images of RT–PCR products from 5 bladders for each of the three groups. GAPDH was used as an internal control. M. A 100-bp DNA ladder. The image was cropped from five different gels run and exposed under the same conditions. Bottom panel: RT-PCR bands were quantified by densitometry and normalized to Gapdh. Data shown are the mean ± SD relative gene expressions. ∗p < 0.01 and ∗∗p < 0.001. (D) Changes in pelvic nociceptive responses during treatment. The data shown are the mean ± SD sensory threshold for 8 baseline mice, 7 mIFNγ6-1mh-treated cystitis mice, 5 PBS-treated cystitis mice and 5 cystitis-uninduced mice. ∗p < 0.05 and ∗∗p < 0.01. (E) Changes in urinary frequency during treatment. The data shown are the mean ± SE urinary frequency for 8 baseline mice and 4–5 mIFNγ6-1mh-treated cystitis mice, PBS-treated cystitis mice, and cystitis-uninduced mice. ∗p < 0.05. See also Figures S4 and S5.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Homma Y., Akiyama Y., Tomoe H., Furuta A., Ueda T., Maeda D., Lin A.T., Kuo H.C., Lee M.H., Oh S.J., et al. Clinical guidelines for interstitial cystitis/bladder pain syndrome. Int. J. Urol. 2020;27:578–589. doi: 10.1111/iju.14234. - DOI - PubMed
    1. Clemens J.Q., Erickson D.R., Varela N.P., Lai H.H. Diagnosis and Treatment of Interstitial Cystitis/Bladder Pain Syndrome. J. Urol. 2022;208:34–42. doi: 10.1097/JU.0000000000002756. - DOI - PubMed
    1. Patnaik S.S., Laganà A.S., Vitale S.G., Butticè S., Noventa M., Gizzo S., Valenti G., Rapisarda A.M.C., La Rosa V.L., Magno C., et al. Etiology, pathophysiology and biomarkers of interstitial cystitis/painful bladder syndrome. Arch. Gynecol. Obstet. 2017;295:1341–1359. doi: 10.1007/s00404-017-4364-2. - DOI - PubMed
    1. Fall M., Nordling J., Cervigni M., Dinis Oliveira P., Fariello J., Hanno P., Kåbjörn-Gustafsson C., Logadottir Y., Meijlink J., Mishra N., et al. Hunner lesion disease differs in diagnosis, treatment and outcome from bladder pain syndrome: an ESSIC working group report. Scand. J. Urol. 2020;54:91–98. doi: 10.1080/21681805.2020.1730948. - DOI - PubMed
    1. Akiyama Y., Maeda D., Katoh H., Morikawa T., Niimi A., Nomiya A., Sato Y., Kawai T., Goto A., Fujimura T., et al. Molecular Taxonomy of Interstitial Cystitis/Bladder Pain Syndrome Based on Whole Transcriptome Profiling by Next-Generation RNA Sequencing of Bladder Mucosal Biopsies. J. Urol. 2019;202:290–300. doi: 10.1097/JU.0000000000000234. - DOI - PubMed