TREATMENT AND PREVENTION OF DISEASE CAUSED BY TYPE IV COLLAGEN DYSFUNCTION
20230212279 · 2023-07-06
Assignee
Inventors
Cpc classification
C07K2317/76
CHEMISTRY; METALLURGY
International classification
Abstract
Methods of treating and preventing Alport syndrome through inhibiting interleukin 11 (EL-11)-mediated signalling are disclosed, as well as agents for use in such methods.
Claims
1. An agent capable of inhibiting interleukin 11 (IL-11)-mediated signalling for use in a method of treating or preventing Alport syndrome.
2. Use of an agent capable of inhibiting interleukin 11 (IL-11)-mediated signalling in the manufacture of a medicament for use in a method of treating or preventing Alport syndrome.
3. A method of treating or preventing Alport syndrome, comprising administering a therapeutically or prophylactically effective amount of an agent capable of inhibiting interleukin 11 (IL-11)-mediated signalling to a subject.
4. The agent for use according to claim 1, the use according to claim 2, or the method according to claim 3, wherein the agent is an agent capable of preventing or reducing the binding of interleukin 11 (IL-11) to a receptor for interleukin 11 (IL-11R).
5. The agent for use according to claim 1 or claim 4, the use according to claim 2 or claim 4, or the method according to claim 3 or claim 4, wherein the agent is capable of binding to interleukin 11 (IL-11) or a receptor for interleukin 11 (IL-11R).
6. The agent for use according to any one of claim 1, 4 or 5, the use according to any one of claim 2, 4 or 5, or the method according to any one of claims 3 to 5, wherein the agent is selected from the group consisting of: an antibody or an antigen-binding fragment thereof, a polypeptide, a peptide, a nucleic acid, an oligonucleotide, an aptamer or a small molecule.
7. The agent for use, the use or the method according to claim 5 or claim 6, wherein the agent is an antibody or an antigen-binding fragment thereof.
8. The agent for use, the use or the method according to claim 7, wherein the agent is an anti-IL-11 antibody antagonist of IL-11-mediated signalling, or an antigen-binding fragment thereof.
9. The agent for use, the use or the method according to claim 7 or claim 8, wherein the antibody or antigen-binding fragment comprises: (i) a heavy chain variable (VH) region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO:34 HC-CDR2 having the amino acid sequence of SEQ ID NO:35 HC-CDR3 having the amino acid sequence of SEQ ID NO:36; and (ii) a light chain variable (VL) region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:37 LC-CDR2 having the amino acid sequence of SEQ ID NO:38 LC-CDR3 having the amino acid sequence of SEQ ID NO:39.
10. The agent for use, the use or the method according to claim 7 or claim 8, wherein the antibody or antigen-binding fragment comprises: (i) a heavy chain variable (VH) region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO:40 HC-CDR2 having the amino acid sequence of SEQ ID NO:41 HC-CDR3 having the amino acid sequence of SEQ ID NO:42; and (ii) a light chain variable (VL) region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:43 LC-CDR2 having the amino acid sequence of SEQ ID NO:44 LC-CDR3 having the amino acid sequence of SEQ ID NO:45.
11. The agent for use, the use or the method according to claim 7, wherein the agent is an anti-IL-11Rα antibody antagonist of IL-11-mediated signalling, or an antigen-binding fragment thereof.
12. The agent for use, the use or the method according to claim 7 or claim 11, wherein the antibody or antigen-binding fragment comprises: (i) a heavy chain variable (VH) region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO:46 HC-CDR2 having the amino acid sequence of SEQ ID NO:47 HC-CDR3 having the amino acid sequence of SEQ ID NO:48; and (ii) a light chain variable (VL) region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:49 LC-CDR2 having the amino acid sequence of SEQ ID NO:50 LC-CDR3 having the amino acid sequence of SEQ ID NO:51.
13. The agent for use, the use or the method according to claim 5 or claim 6, wherein the agent is a decoy receptor.
14. The agent for use, the use or the method according to claim 13, wherein the agent is a decoy receptor for IL-11.
15. The agent for use, the use or the method according to claim 14, wherein the decoy receptor for IL-11 comprises: (i) an amino acid sequence corresponding to the cytokine binding module of gp130 and (ii) an amino acid sequence corresponding to the cytokine binding module of IL-11Rα.
16. The agent for use, the use or the method according to claim 5 or claim 6, wherein the agent is an IL-11 mutein.
17. The agent for use, the use or the method according to claim 16, wherein the IL-11 mutein is W147A.
18. The agent for use according to claim 1, the use according to claim 2, or the method according to claim 3, wherein the agent is capable of preventing or reducing the expression of interleukin 11 (IL-11) or a receptor for interleukin 11 (1L-11R).
19. The agent for use, the use, or the method according to claim 18, wherein the agent is an oligonucleotide or a small molecule.
20. The agent for use, the use or the method according to claim 19, wherein the agent is an antisense oligonucleotide capable of preventing or reducing the expression of IL-11.
21. The agent for use, the use or the method according to claim 20, wherein the antisense oligonucleotide capable of preventing or reducing the expression of IL-11 is siRNA targeted to IL11 comprising the sequence of SEQ ID NO:12, 13, 14 or 15.
22. The agent for use, the use or the method according to claim 19, wherein the agent is an antisense oligonucleotide capable of preventing or reducing the expression of IL-11Rα.
23. The agent for use, the use or the method according to claim 22, wherein the antisense oligonucleotide capable of preventing or reducing the expression of IL-11Rα is siRNA targeted to IL11RA comprising the sequence of SEQ ID NO:16, 17, 18 or 19.
24. The agent for use, the use or the method according to any one of claims 4 to 23, wherein the interleukin 11 receptor is or comprises IL-11Rα.
25. The agent for use according to any one of claims 1, or 4 to 24, the use according to any one of claims 2, or 4 to 24, or the method according to any one of claims 3 to 24, wherein the method comprises administering the agent to a subject in which expression of interleukin 11 (1L-11) or a receptor for IL-11 (1L-11R) is upregulated.
26. The agent for use according to any one of claims 1, or 4 to 25, the use according to any one of claims 2, or 4 to 25, or the method according to any one of claims 3 to 25, wherein the method comprises administering the agent to a subject in expression of interleukin 11 (1L-11) or a receptor for interleukin 11 (1L-11R) has been determined to be upregulated.
27. The agent for use according to any one of claims 1, or 4 to 26, the use according to any one of claims 2, or 4 to 26, or the method according to any one of claims 3 to 26, wherein the method comprises determining whether expression of interleukin 11 (1L-11) or a receptor for IL-11 (1L-11R) is upregulated in the subject and administering the agent to a subject in which expression of interleukin 11 (1L-11) or a receptor for IL-11 (1L-11R) is upregulated.
Description
BRIEF DESCRIPTION OF THE FIGURES
[0493] Embodiments and experiments illustrating the principles of the invention will now be discussed with reference to the accompanying figures.
[0494]
[0495]
[0496]
[0497]
[0498]
EXAMPLES
[0499] In the following Examples, the inventors demonstrate that inhibition of IL-11-mediated signalling reduces increases survival of subjects in a model of Alport syndrome.
[0500] The inventors show that the fibro-inflammatory cytokine interleukin 11 (IL11) is upregulated in the kidneys of Col4a3.sup.−/− mice, and that the receptor for IL11 (IL11RA1) is expressed on podocytes and tubule cells. Administering Col4a3.sup.−/− mice with neutralizing IL11 antibody (X203) at 6-weeks of age (a time when ACE inhibition is no longer effective in slowing progression of kidney disease in this model) is shown to reduce kidney fibrosis, inflammation and tubule damage, improve kidney function and extend lifespan by 41.6%. Given the excellent translatability of Col4a3.sup.−/− mouse as a model of Alport syndrome, the data indicate that IL11-targeted therapies are useful for the treatment of Alport syndrome in humans.
Example 1: Analysis of the Effect of Antagonism of IL-11-Mediated Signalling In Vivo in a Mouse Model of Alport Syndrome
[0501] The inventors investigated the effect of administration of an antagonist of IL-11-mediated signalling on survival of subjects in a mouse model of Alport syndrome.
[0502] 129-Col4a3.sup.tm1Dec/J mice were obtained from The Jackson Laboratory (Stock No:002908; COL4A3 KO). The mice comprise the Co/4a3.sup.tm1Dec mutation described e.g. in Cosgrove et al., Genes Dev. (1996) 10:2981-2992, which is hereby incorporated by reference in its entirety. 129-Col4a3.sup.tm1Dec/J mice comprise a targeted mutation to Col4a3, resulting in progressive glomerulonephritis with microhematuria and proteinuria, and mice homozygous for the mutation typically die at about 8.5 weeks.
[0503] From 6 weeks of age, mice homozygous for the Co/4a3.sup.tm1Dec mutation were administered biweekly by intraperitoneal injection of 20 mg/kg of Enx203, or an isotype-matched control antibody, until mortality.
[0504] Enx203 is a mouse anti-mouse IL-11 IgG, and is described e.g. in Ng et al., Sci Transl Med. (2019) 11(511) pii: eaaw1237 (also published as Ng, et al., “IL-11 is a therapeutic target in idiopathic pulmonary fibrosis.” bioRxiv 336537; doi: https://doi.orq/10.1101/336537). Enx203 is also referred to as “X203”. Enx203 comprises the VH region according to SEQ ID NO:92 of WO 2019/238882 A1 (SEQ ID NO:22 of the present disclosure), and the VL region according to SEQ ID NO:94 of WO 2019/238882 Al (SEQ ID NO:23 of the present disclosure).
[0505] The results of the experiment are shown in
[0506] Thus the inventors establish antagonism of IL-11-mediated signalling as a useful therapeutic intervention for Alport syndrome.
[0507] This is the first therapeutic intervention found to be effective to increase survival of mice in this model when commenced later than 4 weeks after birth.
Example 2: A Neutralising Antibody Antagonist of IL-11-Mediated Signalling Improves Renal Function and Increases Lifespan in a Mouse Model of Alport Syndrome
[0508] 2.1 Overview
[0509] Background: Alport syndrome is a genetic disorder characterized by a defective glomerular basement membrane, tubulointerstitial fibrosis and progressive renal failure. The role of interleukin 11 (IL11) in Alport syndrome is unknown.
[0510] Methods: The effects of a neutralizing IL11 antibody (X203) were assessed, as compared to IgG control, in Col4a3.sup.−/− mice, from six weeks of age, on lifespan, renal tubule damage, function, fibrosis, and inflammation using histology, qPCR, immunoblotting and immunohistochemistry.
[0511] Results: Renal expression of IL11 is elevated in Col4a3.sup.−/− mice and the IL11RA1 receptor is found predominantly in tubular cells and podocytes. Administration of X203 reduced albuminuria and improved renal function, assessed by BUN and serum creatinine levels. X203 also extended the median life span of Col4a3.sup.−/− mice by 41.6%, from 62.5 to 88.5 days. Podocyte numbers and levels of key podocytes proteins including Wilms' Tumor 1 and Podocin, which are reduced in Col4a3.sup.−/− mice, were restored towards normal by X203 administration. The beneficial effects of X203 on kidney structure and function were accompanied by reduced severity of renal fibrosis and inflammation, markers of tubule damage, and features of epithelial-to-mesenchymal transition. Pathogenic ERK and STAT3 activities were elevated in Col4a3.sup.−/− mice and these pathways were largely reduced by administration of X203.
[0512] Conclusion: In a mouse model of Alport syndrome, IL11 is upregulated in the kidney and a neutralizing IL11 antibody, given at a time point when angiotensin-converting enzyme inhibition is ineffective, improves kidney structure and function while extending lifespan.
[0513] 2.2 Materials and Methods
[0514] Antibodies
[0515] ACTA2 (19245, CST; WB), Cyclin D1 (55506, CST), E-Cadherin (3195, CST), p-ERK1/2 (4370, CST), ERK1/2 (4695, CST), GAPDH (2118, CST), IgG (11E10, Aldevron; which is produced from 1.10E+11 cells (ATCC, No. CRL-1907)), neutralizing anti-IL11 (X203, Aldevron), anti-IL11RA (X209, Aldevron), NHPS2/Podocin (ab181143, Abcam), PCNA (13110, CST), SNAIL (3879, CST, WB), p-STAT3 (4113, CST), STAT3 (4904, CST), TGFβ (3711, CST), Wilms' Tumor 1 (ab89901, Abcam, IHC, Wilms' Tumor 1 (ab267377, Abcam, WB), anti-rabbit HRP (7074, CST), anti-mouse HRP (7076, CST).
[0516] Mouse Model of Alport
[0517] Animal studies were carried out in compliance with the recommendations in the Guidelines on the Care and Use of Animals for Scientific Purposes of the National Advisory Committee for Laboratory Animal Research (NACLAR). All experimental procedures were approved (SHS/2019/1482) and conducted in accordance with the SingHealth Institutional Animal Care and Use Committee.
[0518] Col4a3.sup.−/− (Col4a3.sup.tm1Dec) mice were purchased from The Jackson Laboratory (https://www.jax.org/strain/002908). Mice were housed in temperatures of 21-24° C. with 40-70% humidity on a 12 h light/12 h dark cycle and provided with food and water ad libitum. For treatment study, Col4a3.sup.−/− were administered 20 mg/kg of anti-IL11 (X203) or IgG isotype control (11E10) by intraperitoneal (IP) injection starting from 6 weeks of age twice a week for 2.5 weeks; wild-type littermates were used as controls. Mice were sacrificed for blood and kidney collection when they were 8.5-week-old. For lifespan study, IgG/X203 was administered to 6-week-old Col4a3.sup.−/− (2×/week) until death ensued.
[0519] Western Blot
[0520] Western blot was carried out on total protein extracts from mouse kidney tissues. Kidneys were lysed in radioimmunoprecipitation assay (RIPA) buffer containing protease and phosphatase inhibitors (Thermo Scientifics), followed by centrifugation to clear the lysate. Protein concentrations were determined by Bradford assay (Bio-Rad). Protein lysates were separated by SDS-PAGE, transferred to PVDF membrane, and subjected to immunoblot analysis for various antibodies as outlined in the main text, figures, or and/or figure legends. Proteins were visualized using the ECL detection system (Pierce) with the appropriate secondary antibodies: anti-rabbit HRP or antimouse HRP.
[0521] Quantitative Polymerase Chain Reaction (qPCR)
[0522] Total RNA was extracted from snap-frozen kidney tissues using Trizol (Invitrogen) followed by RNeasy column (Qiagen) purification. cDNAs were synthesized with iScript™ cDNA synthesis kit (Bio-Rad) according to manufacturer's instructions. Gene expression analysis was performed on duplicate samples with either TaqMan (Applied Biosystems) or fast SYBR green (Qiagen) technology using StepOnePlus™ (Applied Biosystem) over 40 cycles. Expression data were normalized to GAPDH mRNA expression and fold change was calculated using 2.sup.−ΔΔct method. The sequences of specific TaqMan probes and SYBR green primers are available upon request.
[0523] Colorimetric Assays
[0524] The levels of blood urea nitrogen (BUN) and creatinine in mouse serum were measured using Urea Assay Kit (ab83362, Abcam) and Creatinine Assay Kit (ab65340, Abcam), respectively. Urine albumin and creatinine levels were measured using Mouse Albumin ELISA kit (ab108792, Abcam) and Creatinine Assay Kit (ab204537, Abcam), respectively. All ELISA and colorimetric assays were performed according to the manufacturer's protocol.
[0525] Masson Trichrome's Staining
[0526] Kidney tissues were fixed for 48 hours at RT in 10% neutral-buffered formalin (NBF), dehydrated, embedded in paraffin, and sectioned at 7 μm. Transverse kidney sections were then stained with Masson's Trichrome according to standard protocol. Images of the sections were captured by light microscopy and blue-stained fibrotic areas were semi-quantitatively determined with Image-J software (color deconvolution-Masson Trichrome) from the whole kidney area (100× field, n=4 kidneys/group). Treatment and genotypes were not disclosed to investigators performing the histology and generating semi-quantitative readouts.
[0527] Immunohistochemistry
[0528] Kidneys were fixed in 10% neutral-buffered formalin (NBF), paraffinized, cut into 7 μm sections, incubated with primary antibodies overnight and visualized using an ImmPRESS HRP anti-rabbit IgG polymer detection kit (MP-7401, Vector Laboratories) with ImmPACT DAB Peroxidase Substrate (SK-4105, Vector Laboratories). Quantification of WT.sup.+ve cells were performed in a blinded fashion from 4 images (200× field)/kidney (n=3-4 kidneys/group).
[0529] Statistical Analyses
[0530] Statistical analyses were performed using GraphPad Prism software (version 8). Statistical significance between control and experimental groups were analysed by twosided Student's t tests or by one-way ANOVA as indicated in the figure legends. P values were corrected for multiple testing according to Tukey when several conditions were compared to each other within one experiment. Comparison analysis for two parameters from two different groups were performed by two-way ANOVA. Survival curves were analyzed by Gehan-Breslow-Wilcoxon test. The criterion for statistical significance was P<0.05.
[0531] 2.3 Results
[0532] IL11 is Upregulated in the Kidneys of Col4a3.sup.−/− Mice.
[0533] IL11 is not expressed in normal healthy tissues but its induction is commonly seen in fibroinflammatory diseases.sup.15. Expression of II11 mRNA was profiled in kidneys of Col4a3.sup.−/− mice, and was found to be upregulated (17.8-fold, P<0.0001), as compared to wild-type littermate controls (
[0534] Stromal cells, epithelial cells, and other cells can express IL11, and so the cells in the kidney that express the IL11 receptor (IL11RA1) were determined by both immunohistochemistry and mining publicly available single cell RNA sequencing (scRNA-seq) data.sup.16. In wild-type mice, IL11RA1 expression was easily seen in tubules and also in the glomerulus, whereas no staining was seen in sections from II11ra1.sup.−/− mice, confirming specificity of detection (
[0535] Antibody Neutralization of IL11 Reduces Molecular Pathologies in Col4a3.sup.−/− Mice.
[0536] Over recent years, antibodies that inhibit IL11 signaling in mouse and human cells have been developed.sup.16,17. One of these neutralizing IL11 antibodies (X203) or an IgG control were administered to 6-week-old Col4a3.sup.−/− mice, the time point when initiation of ramipril has proven ineffective.sup.6, and examined renal pathologies at 8.5 weeks of age, as compared to age-matched wild-type controls (
[0537] At the end of the study period, total body weight loss, measured as a percentage of their starting body weight, was significantly attenuated in Col4a3.sup.−/− mice receiving X203, compared to those administered with IgG (IgG: 30%, X203:17%; P=0.0003) (
[0538] At the signaling level, IL11 is known to activate ERK across primary cell types and this pathway has been mechanistically linked with MI-driven fibrosis.sup.17-19. IL11 inhibition in vivo can also be associated with reduced STAT3 activation, which is thought to be a secondary phenomenon reflecting lesser stromal-driven inflammation.sup.17,20,21. As compared to wild-type mice, kidneys from Col4a3.sup.−/− mice treated with IgG exhibited elevated ERK and STAT3 activation, in contrast ERK and STAT3 phosphorylation was largely diminished in kidneys of X203-treated Col4a3.sup.−/− mice (
[0539] In many kidney diseases, it is thought that damaged TECs undergo a pEMT process, which is critical for the subsequent development of tubulointerstitial fibrosis and CKD.sup.22,23. TEC pEMT is characterised by increased SNAI1 expression and reciprocal downregulation of E-Cadherin, which is regulated in part by TGFβ22,23. As compared to wild-type controls, Col4a3.sup.−/− mice receiving IgG exhibited a strong molecular signature of EMT with increased SNAI1 and decreased E-Cadherin expression (
[0540] Podocyte Preservation and Lesser Renal Inflammation is Associated with Inhibition of IL11 Signaling in Col4a3.sup.−/− Mice.
[0541] AS affects GBM composition leading to podocyte dysfunction/loss that relates to TGFβ activity in both podocytes and TECs.sup.9. Immunohistochemistry analysis of the podocyte marker, Wilms' Tumor 1 (WT1) revealed a greater staining in wild-type mice and X203-treated Col4a3.sup.−/− mice, as compared to IgG-treated Col4a3.sup.−/− mice (
[0542] TGFβ upregulation in podocytes and tubular cells, which is coincident with the onset of proteinuria in the Col4a3.sup.−/− mouse.sup.9,24, is thought of importance for disease pathogenesis in AS. Levels of TGFβ were therefore examined, and it was observed that X203, but not IgG, significantly reduced the degree of TGFβ upregulation in the kidneys of Col4a3.sup.−/− mice (
[0543] Tnfα expression in podocytes is of particular importance in AS and leads to podocyte apoptosis and glomerulosclerosis.sup.13. It was therefore notable that X203 reduced Tnfα expression in Col4a3.sup.−/− mice, as compared to IgG treated controls (
[0544] Inhibition of IL11 Signaling Improves Renal Function and Prolongs Lifespan in Col4a3.sup.−/− Mice.
[0545] Next, it was investigated whether inhibition of IL11 signaling, which mitigated intermediate phenotypes of renal pathology in Col4a3.sup.−/− mice, also improved renal function. To do so serum blood urea nitrogen (BUN) and creatinine (Cr) and also urinary albumin:creatinine ratios were determined at study end (8.5 weeks of age). As compared to wild-type mice, IgG-treated Col4a3.sup.−/− mice had elevated BUN, Cr and urinary ACR levels (fold elevation compared to wild-type: 12.4, 7.3, 13.6, respectively), whereas administration of X203 from week 6 markedly lowered BUN, Cr and urinary ACR consistent with an overall improvement in renal function (
[0546] Death from progressive kidney failure is consistently seen in untreated Col4a3.sup.−/− mice from 8.5 weeks of age and mean survival times are reproducibly reported at around 10 weeks (e.g. 71 days.sup.6; 69 days.sup.26). The Col4a3.sup.−/− mice used for the studies described here had a similar mean survival of 62.7±1.9 days, albeit with a trend towards a shorter lifespan. It was investigated whether administration of X203 (20 mg/kg; 2×/week) to 6-week old Col4a3.sup.−/− mice extended life, as compared to IgG control-treated mice (
[0547] 2.4 Discussion
[0548] While ACEi is the mainstay of therapy for patients with AS, progression to endstage renal failure is typical.sup.6,10. This shortcoming likely reflects the complex renal pathology in AS, involving GBM-specific initiating factors and generic tubulointerstitial disease mechanisms. ACEi may impact a number of AS pathologies and reducing ultrafiltration is thought of as of primary importance. As IL11 antibody therapy is beneficial at 6 weeks of age in Col4a3.sup.−/− mice, when ACEi is ineffective, this suggests an alternative mechanism of action is associated with inhibition of IL11.
[0549] IL11 is secreted from a variety of stromal and epithelial cells in response to cellular injury and acts in an autocrine and paracrine manner to cause epithelial cell dysfunction, stromal activation and tissue inflammationm. In the kidney, IL11RA is expressed on tubule cells throughout the nephron and in podocytes, two cell types that can be affected by pEMT.sup.22,23,26. One way that inhibition of IL11 signaling is protective in Col4a3.sup.−/− mice could be through inhibition of pEMT, thus preserving both podocyte and TEC function. This could be secondary to lower TGFβ, a master determinant of pEMT in the kidney.sup.22,23, after X203 administration. While inhibition of TGFβ directly is proinflammatory.sup.27 it is shown here in the kidney, as in other tissues, that inhibition of IL11 is anti-inflammatory.sup.17,21,28.
[0550] The results suggest that inhibition of IL11 signaling is a plausible therapeutic approach for patients with AS and perhaps other forms of progressive CKD, especially as new IL11-targeting therapies are progressing towards the clinic. Besides limited developmental bone/tooth defects, humans with loss-of-function in IL11RA are otherwise well with normal immune function and lifespans, as are II11ra1 null mice. Furthermore, two recently and separately developed II11 null mice appear normal with no bone deficits.sup.29,30 and it could be that inhibiting the IL11 cytokine has advantages over targeting IL11RA. Taken together, the human and mouse 1L11RA and II11 genetic null data, along with long-term antibody studies in mice.sup.17, provide a safety signal needed to consider long term trials of IL11 inhibition in AS.sup.16,31.
REFERENCES
[0551] 1. Hudson B G, Tryggvason K, Sundaramoorthy M, Neilson E G: Alport's Syndrome, Goodpasture's Syndrome, and Type IV Collagen [Internet]. New England Journal of Medicine. 348: 2543-2556, 2003 Available from: http://dx.doi.org/10.1056/nejmra022296
[0552] 2. Hudson B G, Reeders S T, Tryggvason K: Type IV collagen: structure, gene organization, and role in human diseases. Molecular basis of Goodpasture and Alport syndromes and diffuse leiomyomatosis. J. Biol. Chem. 268: 26033-26036, 1993 [0553] 3. Savige J: Alport syndrome: its effects on the glomerular filtration barrier and implications for future treatment. J. Physiol. 592: 4013-4023, 2014 [0554] 4. Funk S D, Lin M-H, Miner J H: Alport syndrome and Pierson syndrome: Diseases of the glomerular basement membrane. Matrix Biol. 71-72: 250-261, 2018 [0555] 5. Jais J P, Knebelmann B, Giatras I, De Marchi M, Rizzoni G, Renieri A, Weber M, Gross O, Netzer K O, Flinter F, Pirson Y, Verellen C, Wieslander J, Persson U, Tryggvason K, Martin P, Hertz J M, Schröder C, Sanak M, Krejcova S, Carvalho M F, Saus J, Antignac C, Smeets H, Gubler M C: X-linked Alport syndrome: natural history in 195 families and genotype-phenotype correlations in males. J. Am. Soc. Nephrol. 11: 649-657, 2000 [0556] 6. Gross O, Beirowski B, Koepke M-L, Kuck J, Reiner M, Addicks K, Smyth N, Schulze-Lohoff E, Weber M: Preemptive ramipril therapy delays renal failure and reduces renal fibrosis in COL4A3-knockout mice with Alport syndrome. Kidney Int 63: 438-446, 2003 [0557] 7. Gross O, Tönshoff B, Weber L T, Pape L, Latta K, Fehrenbach H, Lange-Sperandio B, Zappel H, Hoyer P, Staude H, König S, John U, Gellermann J, Hoppe B, Galiano M, Hoecker B, Ehren R, Lerch C, Kashtan C E, Harden M, Boeckhaus J, Friede T, German Pediatric Nephrology (GPN) Study Group and EARLY PRO-TECT Alport Investigators: A multicenter, randomized, placebo-controlled, double-blind phase 3 trial with open-arm comparison indicates safety and efficacy of nephroprotective therapy with ramipril in children with Alport's syndrome. Kidney Int. 97: 1275-1286, 2020 [0558] 8. Gross O, Licht C, Anders H J, Hoppe B, Beck B, Tonshoff B, Hocker B, Wygoda S, Ehrich J H H, Pape L, Konrad M, Rascher W, Dotsch J, Müller-Wiefel D E, Hoyer P, Study Group Members of the Gesellschaft für Pädiatrische Nephrologie, Knebelmann B, Pirson Y, Grunfeld J-P, Niaudet P, Cochat P, Heidet L, Lebbah S, Torra R, Friede T, Lange K, Müller G A, Weber M: Early angiotensin-converting enzyme inhibition in Alport syndrome delays renal failure and improves life expectancy. Kidney Int 81: 494-501, 2012 [0559] 9. Sayers R, Kalluri R, Rodgers K D, Shield C F, Meehan D T, Cosgrove D: Role for transforming growth factor-beta1 in alport renal disease progression. Kidney Int 56: 1662-1673, 1999 [0560] 10. Kashtan C E: Renal transplantation in patients with Alport syndrome: patient selection, outcomes, and donor evaluation. Int. J. Nephroi. Renovasc. Dis. 11: 267-270, 2018 [0561] 11. Hood J C, Dowling J, Bertram J F, Young R J, Huxtable C, Robinson W, Savige J: Correlation of histopathological features and renal impairment in autosomal dominant Alport syndrome in Bull terriers. Nephrol. Dial. Transplant 17: 1897-1908, 2002 [0562] 12. Qi R, Yang C: Renal tubular epithelial cells: the neglected mediator of tubulointerstitial fibrosis after injury. Cell Death Dis. 9: 1126, 2018 [0563] 13. Ryu M, Mulay S R, Miosge N, Gross O, Anders H-J: Tumour necrosis factor-α drives Alport glomerulosclerosis in mice by promoting podocyte apoptosis. J. Pathol. 226: 120-131, 2012 [0564] 14. Schafer S, Viswanathan S, Widjaja A A, Lim W-W, Moreno-Moral A, DeLaughter D M, Ng B, Patone G, Chow K, Khin E, Tan J, Chothani S P, Ye L, Rackham O J L, Ko NSJ, Sahib N E, Pua C J, Zhen N T G, Xie C, Wang M, Maatz H, Lim S, Saar K, Blachut S, Petretto E, Schmidt S, Putoczki T, Guimarães-Camboa N, Wakimoto H, van Heesch S, Sigmundsson K, Lim S L, Soon J L, Chao V T T, Chua Y L, Tan T E, Evans S M, Loh Y J, Jamal M H, Ong K K, Chua K C, Ong B-H, Chakaramakkil M J, Seidman J G, Seidman C E, Hubner N, Sin K Y K, Cook S A: IL-11 is a crucial determinant of cardiovascular fibrosis. Nature 552: 110-115, 2017 [0565] 15. Cook S A, Schafer S: Hiding in Plain Sight: Interleukin-11 Emerges as a Master Regulator of Fibrosis, Tissue Integrity, and Stromal Inflammation. Annu. Rev. Med. 71: 263-276, 2020 [0566] 16. Park J, Shrestha R, Qiu C, Kondo A, Huang S, Werth M, Li M, Barasch J, Susztak K: Single-cell transcriptomics of the mouse kidney reveals potential cellular targets of kidney disease. Science [Internet] 2018 Available from: http://dx.doi.org/10.1126/science.aar2131 [0567] 17. Widjaja A A, Singh B K, Adami E, Viswanathan S, Dong J, D'Agostino G A, Ng B, Lim W W, Tan J, Paleja B S, Tripathi M, Lim S Y, Shekeran S G, Chothani S P, Rabes A, Sombetzki M, Bruinstroop E, Min L P, Sinha R A, Albani S, Yen P M, Schafer S, Cook S A: Inhibiting Interleukin 11 Signaling Reduces Hepatocyte Death and Liver Fibrosis, Inflammation, and Steatosis in Mouse Models of Nonalcoholic Steatohepatitis. Gastroenterology 157: 777-792.e14, 2019 [0568] 18. Lim W-W, Corden B, Ye L, Viswanathan S, Widjaja A A, Xie C, Su L, Tee N G Z, Schafer S, Cook S A: Antibody-mediated neutralization of IL11 signalling reduces ERK activation and cardiac fibrosis in a mouse model of severe pressure overload. Clin. Exp. Pharmacol. Physiol. [Internet] 2021 Available from: http://dx.doi.org/10.1111/1440-1681.13458 [0569] 19. Adami E, Viswanathan S, Widjaja A A, Ng B, Chothani S, Zhihao N, Tan J, Lio P M, George B L, Altunoglu U, Ghosh K, Paleja B S, Schafer S, Reversade B, Albani S, Ling A L H, O'Reilly S, Cook S A: IL11 is elevated in systemic sclerosis and IL11-dependent ERK signaling underlies TGFβ-mediated activation of dermal fibroblasts. Rheumatology [Internet] 2021 Available from: http://dx.doi.org/10.1093/rheumatology/keab168 [0570] 20. Dong J, Viswanathan S, Adami E, Singh B K, Chothani S P, Ng B, Lim V W V, Zhou J, Tripathi M, Ko N S J, Shekeran S G, Tan J, Lim S Y, Wang M, Lio P M, Yen P M, Schafer S, Cook S A, Widjaja A A: Hepatocyte-specific IL11 cis-signaling drives lipotoxicity and underlies the transition from NAFLD to NASH. Nat. Commun. 12: 66, 2021 [0571] 21. Ng B, Dong J, Viswanathan S, Widjaja A A, Paleja B S, Adami E, Ko N S J, Wang M, Lim S, Tan J, Chothani S P, Albani S, Schafer S, Cook S A: Fibroblast-specific IL11 signaling drives chronic inflammation in murine fibrotic lung disease. FASEB J. [Internet] 2020 Available from: http://dx.doi.org/10.1096/fj.202001045RR [0572] 22. Lovisa S, LeBleu V S, Tampe B, Sugimoto H, Vadnagara K, Carstens J L, Wu C-C, Hagos Y, Burckhardt B C, Pentcheva-Hoang T, Nischal H, Allison J P, Zeisberg M, Kalluri R: Epithelial-to-mesenchymal transition induces cell cycle arrest and parenchymal damage in renal fibrosis. Nat. Med. 21: 998-1009, 2015 [0573] 23. Grande M T, Sanchez-Laorden B, Lopez-Blau C, De Frutos C A, Boutet A, Arevalo M, Rowe R G, Weiss S J, Lopez-Novoa J M, Nieto M A: Snaill-induced partial epithelial-to-mesenchymal transition drives renal fibrosis in mice and can be targeted to reverse established disease. Nat. Med. 21: 989-997, 2015 [0574] 24. Suh S H, Choi H S, Kim C S, Kim I J, Ma S K, Scholey J W, Kim S W, Bae E H: Olmesartan Attenuates Kidney Fibrosis in a Murine Model of Alport Syndrome by Suppressing Tubular Expression of TGFβ. Int. J. Mol. Sci. [Internet] 20: 2019 Available from: http://dx.doi.org/10.3390/ijms20153843 [0575] 25. Ninichuk V, Gross O, Reichel C, Khandoga A, Pawar R D, Ciubar R, Segerer S, Belemezova E, Radomska E, Luckow B, Perez de Lema G, Murphy P M, Gao J-L, Henger A, Kretzler M, Horuk R, Weber M, Krombach F, Schlondörff D, Anders H-J: Delayed chemokine receptor 1 blockade prolongs survival in collagen 4A3-deficient mice with Alport disease. J. Am. Soc. Nephroi. 16: 977-985, 2005 [0576] 26. Ying Q, Wu G: Molecular mechanisms involved in podocyte EMT and concomitant diabetic kidney diseases: an update. Ren. Fail. 39: 474-483, 2017 27. Shull M M, Ormsby I, Kier A B, Pawlowski S, Diebold R J, Yin M, Allen R, Sidman C, Proetzel G, Calvin D: Targeted disruption of the mouse transforming growth factorbeta 1 gene results in multifocal inflammatory disease. Nature 359: 693-699, 1992 [0577] 28. Lim W-W, Corden B, Ng B, Vanezis K, D'Agostino G, Widjaja A A, Song W-H, Xie C, Su L, Kwek X-Y, Tee N G Z, Dong J, Ko N S J, Wang M, Pua C J, Jamal M H, Soh B, Viswanathan S, Schafer S, Cook S A: Interleukin-11 is important for vascular smooth muscle phenotypic switching and aortic inflammation, fibrosis and remodeling in mouse models [Internet]. Scientific Reports. 10: 2020 Available from: http://dx.doi.org/10.1038/s41598-020-74944-7 [0578] 29. Nishina T, Deguchi Y, Ohshima D, Takeda W, Ohtsuka M, Shichino S, Ueha S, Yamazaki S, Kawauchi M, Nakamura E, Nishiyama C, Kojima Y, Adachi-Akahane S, Hasegawa M, Nakayama M, Oshima M, Yagita H, Shibuya K, Mikami T, Inohara N, Matsushima K, Tada N, Nakano H: Interleukin-11-expressing fibroblasts have a unique gene signature correlated with poor prognosis of colorectal cancer. Nat. Commun. 12: 2281, 2021 [0579] 30. Ng B, Widjaja A A, Viswanathan S, Dong J: Similarities and differences between IL11 and IL11RA1 knockout mice for lung fibro-inflammation, fertility and craniosynostosis. bioRxiv [Internet] 2020 Available from: https://www.biorxiv.org/content/10.1101/2020.12.10.420695v1.abstract [0580] 31. Widjaja A A, Chothani S P, Cook S A: Different roles of interleukin 6 and interleukin 11 in the liver: implications for therapy. Hum. Vaccin. Immunother. 16: 2357-2362, 2020