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Sporadic cases of apolipoprotein A-IV medullary amyloidosis have been reported. Here we describe five families found to have autosomal dominant medullary amyloidosis due to two different pathogenic APOA4 variants. A large family with autosomal dominant chronic kidney disease (CKD) and bland urinary sediment underwent whole genome sequencing with identification of a chr11:116692578 G>C (hg19) variant encoding the missense mutation p.L66V of the ApoA4 protein. We identified two other distantly related families from our registry with the same variant and two other distantly related families with a chr11:116693454 C>T (hg19) variant encoding the missense mutation p.D33N. Both mutations are unique to affected families, evolutionarily conserved and predicted to expand the amyloidogenic hotspot in the ApoA4 structure. Clinically affected individuals suffered from CKD with a bland urinary sediment and a mean age for kidney failure of 64.5 years. Genotyping identified 48 genetically affected individuals; 44 individuals had an estimated glomerular filtration rate (eGFR) under 60 ml/min/1.73 m2, including all 25 individuals with kidney failure. Significantly, 11 of 14 genetically unaffected individuals had an eGFR over 60 ml/min/1.73 m2. Fifteen genetically affected individuals presented with higher plasma ApoA4 concentrations. Kidney pathologic specimens from four individuals revealed amyloid deposits limited to the medulla, with the mutated ApoA4 identified by mass-spectrometry as the predominant amyloid constituent in all three available biopsies. Thus, ApoA4 mutations can cause autosomal dominant medullary amyloidosis, with marked amyloid deposition limited to the kidney medulla and presenting with autosomal dominant CKD with a bland urinary sediment. Diagnosis relies on a careful family history, APOA4 sequencing and pathologic studies.
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Amiloidosis , Apolipoproteínas A , Nefritis Intersticial , Insuficiencia Renal Crónica , Humanos , Persona de Mediana Edad , Nefritis Intersticial/diagnóstico , Nefritis Intersticial/genética , Nefritis Intersticial/complicaciones , Mutación , Insuficiencia Renal Crónica/diagnóstico , Insuficiencia Renal Crónica/genética , Insuficiencia Renal Crónica/complicacionesRESUMEN
BACKGROUND AND PURPOSE: Activated fibroblasts deposit fibrotic matrix in chronic kidney disease (CKD) and G-protein coupled receptors (GPCRs) are the most druggable therapeutic targets. Here, we set out to establish a transcriptional profile that identifies activated kidney fibroblasts and the GPCRs that they express. EXPERIMENTAL APPROACH: RNA sequencing and single cell qRT-PCR were performed on mouse kidneys after unilateral ureteral obstruction (UUO). Candidate expression was evaluated in mice with UUO or diabetes or injected with adriamycin or folic acid. Intervention studies were conducted in mice with diabetes or UUO. Correlative histology was performed in human kidney tissue. KEY RESULTS: Transcription factor 21 (Tcf21)+ cells that expressed 2 or 3 of Postn, Acta2 and Pdgfra were highly enriched for fibrogenic genes and were defined as activated kidney fibroblasts. Tcf21+ α-smooth muscle actin (α-SMA)+ interstitial cells accumulated in kidneys of mice with UUO or diabetes or injected with adriamycin or folic acid, whereas renin-angiotensin system blockade attenuated increases in Tcf21 in diabetic mice. Fifty-six GPCRs were up-regulated in single Tcf21+ kidney fibroblasts, the most up-regulated being Adgra2 and S1pr3. Adenosine receptors, Adora2a/2b, were up-regulated in Tcf21+ fibroblasts and the adenosine receptor antagonist, caffeine decreased Tcf21 upregulation and kidney fibrosis in UUO mice. TCF21, ADGRA2, S1PR3 and ADORA2A/2B were each detectable in α-SMA+ interstitial cells in human kidney samples. CONCLUSION AND IMPLICATIONS: Tcf21 is a marker of kidney fibroblasts that are enriched for fibrogenic genes in CKD. Further analysis of the GPCRs expressed by these cells may identify new targets for treating CKD. LINKED ARTICLES: This article is part of a themed issue on Translational Advances in Fibrosis as a Therapeutic Target. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v180.22/issuetoc.
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Diabetes Mellitus Experimental , Enfermedades Renales , Insuficiencia Renal Crónica , Obstrucción Ureteral , Animales , Humanos , Ratones , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Diabetes Mellitus Experimental/metabolismo , Doxorrubicina/farmacología , Fibroblastos/metabolismo , Fibrosis , Ácido Fólico/metabolismo , Ácido Fólico/farmacología , Ácido Fólico/uso terapéutico , Riñón , Enfermedades Renales/metabolismo , Ratones Endogámicos C57BL , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismo , Insuficiencia Renal Crónica/metabolismo , Factores de Transcripción/metabolismo , Obstrucción Ureteral/metabolismoRESUMEN
Introduction: Percutaneous kidney biopsy is the gold standard method to reach a precise diagnosis in most medical kidney diseases, which positively impacts patient care by personalizing the treatment. Accurate diagnosis in the pathology report for medical kidney diseases requires clinicopathological correlation, and clinical data is not always reachable to the nephropathologist. This study aimed to create a standardized, paperless requisition form compatible with medical renal biopsies. Methods: An initial form was prepared for native and allograft renal biopsies according to the current classification of medical kidney diseases. We invited 33 nephropathologists working in Canadian healthcare institutions to answer survey questions about the need to include a particular aspect of clinical information. According to the responses, we modified the experimental form. Eighty nephrologists were asked to complete a clinical data-collecting form given out as PDF files. The time for completing the form and clinicians' satisfaction were assessed. Results: The experimental form survey was answered by 20 out of 33 nephropathologists (61%) from 14 Canadian healthcare centers. The agreement rate on the questions was from 38.89% to 100.00% (average 83.33% and 77.14% for the native and the allograft section, respectively). Seventeen out of 80 nephrologists and their assistants (21%) responded by completing 22 PDF forms. The time required to finish a PDF form was 10.4 min on average. Nephrologists considered the form time-consuming and suggested making it more clinically relevant. Only seven nephrologists responded to the satisfaction survey; four (57%) were satisfied. Conclusions: Medical information is critical in renal pathology diagnoses. A uniform paperless clinical data requisition form was evolved through an agreement by Canadian nephropathologists.
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Objective: To compare the association of margin sampling technique on survival outcomes in surgically treated cT1-2 oral cavity and oropharyngeal squamous cell carcinoma. Study Design: A prospective longitudinal cohort study. Setting: Tertiary care academic teaching hospital in Halifax, Nova Scotia. Methods: All cases of surgically treated cT1-2 oral cavity and oropharyngeal cancer undergoing specimen-oriented margin analysis between January 1, 2017, and December 31, 2018 were analyzed. The specimen-oriented cohort was compared with a cohort of patients from January 1, 2009, to December 31, 2014, where a defect-oriented margin sampling protocol was used. Kaplan-Meier survival curves were used to estimate 2-year overall survival, disease-specific survival, local control, and recurrence-free survival rates in oral cavity and p16-positive oropharyngeal squamous cell carcinoma. Cox proportional hazards models were used to assess the effect of margin sampling method on disease-specific survival and local control. Results: There was no significant association between margin sampling technique and 2-year survival outcomes for surgically treated cT1-2 oral cavity and oropharyngeal squamous cell carcinoma. In the multivariate Cox proportional hazard model, the hazard ratio (HR) of specimen-oriented sampling was not significantly different for disease-specific survival (HR, 1.32; 95% CI, 0.3032-5.727; P = .713) or local control (HR, 0.4087; 95% CI, 0.0795-2.099; P = .284). Conclusion: Intraoperative margin sampling method was not associated with a significant change in 2-year survival outcomes. Despite no effect on survival outcomes, implementation of a specimen-oriented sampling method has potential for cost avoidance by decreasing the number of re-resections for positive or close margins.
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Serum amyloid A protein (AA) amyloidosis, also known as secondary amyloidosis, is a known consequence of chronic inflammation and results from several conditions including inflammatory arthritis, periodic fever syndromes, and chronic infection. AA amyloidosis can lead to multiorgan dysfunction, including changes in glomerular filtration rate and proteinuria. Definitive diagnosis requires tissue biopsy, and management of AA amyloid kidney disease is primarily focused on treating the underlying inflammatory condition to stabilize glomerular filtration rate, reduce proteinuria, and slow potential progression to kidney failure. In this narrative review, we describe the causes, pathophysiology, presentation, and pathologic diagnosis of AA amyloid kidney disease using an illustrative case of biopsy-proven AA amyloid kidney disease in a patient with long-standing rheumatoid arthritis who had a favorable response to interleukin 6 inhibition. We conclude the review with a description of established and more novel therapies for AA amyloidosis including published cases of use of tocilizumab (an interleukin 6 inhibitor) in biopsy-proven AA amyloid kidney disease.
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Rationale: Q fever is a zoonotic infection that may lead to acute or long-term renal injury. Given its rare incidence, Q fever is not often considered on the initial differential diagnosis for glomerular disease which can lead to delays in treatment. This case highlights the importance of avoiding early diagnostic closure and revisiting the differential diagnosis in the setting of an atypical clinical presentation or response to treatment. Presenting Concerns: A 52-year-old female was referred for assessment of possible glomerulonephritis. She described a 3-month history of bilateral lower extremity rash, intermittent knee pain with swelling, and a 2-year history of subjective fevers. Urinalysis showed persistent microscopic hematuria, and her creatinine was elevated at 94 umol/L (baseline 59 umol/L). Her initial investigations included an elevated C-reactive protein (CRP) and rheumatoid factor with a weakly positive anti nuclear antibody (ANA). Diagnoses: Kidney biopsy was consistent with an immune complex mesangial proliferative glomerulonephritis. Light microscopy showed diffuse global mesangial hypercellularity. Immunofluorescence was positive for trace mesangial IgG and kappa, 1+ IgM, lambda and C1q, and 2+ C3. Electron microscopy showed mesangial electron dense deposits. These findings were felt to be most in keeping with mesangial proliferative lupus nephritis; however, it was acknowledged that clinical and laboratory findings supporting this diagnosis were lacking. Interventions: Following treatment with oral prednisone her symptoms resolved, and renal function improved. However, she was unable to taper off prednisone completely without her symptoms returning. Additional immunosuppressive therapies were trialed, but she remained steroid dependent with disease flares related to prednisone tapers. Her atypical response to treatment led to consideration of alternative diagnoses, and further investigation revealed positive Q fever serology (phase-I IgG 1:1892, phase II IgG 1:8192, phase-I and -II IgM < 1:16). She was diagnosed with long-term Q fever and was treated with doxycycline and hydroxychloroquine. Outcomes: She remained on treatment for 2 years. During this time, her symptoms resolved, hematuria disappeared, and her creatinine returned to baseline. Following cessation of therapy, her Q fever IgM titres rose, and she was restarted on doxycycline and hydroxychloroquine indefinitely. Teaching Points: (1) Keeping a broad differential diagnosis in the setting of atypical clinical features or unexpected response to therapy is important for ensuring accurate diagnosis and appropriate treatment. (2) Clinical improvement in relation to immunosuppressive therapy does not preclude an infectious cause of glomerular disease.
Justification: La fièvre Q est une infection zoonotique qui peut entraîner l'insuffisance rénale aiguë ou chronique. D'incidence rare, la fièvre Q n'est généralement pas considérée dans le diagnostic différentiel initial de la glomérulonéphrite, ce qui peut retarder le traitement. Ce cas souligne l'importance d'éviter de poser un diagnostic précoce et d'envisager un diagnostic différentiel lors d'une présentation clinique atypique ou d'une réponse inattendue au traitement. Présentation du cas: Une femme de 52 ans envoyée pour l'évaluation d'une possible glomérulonéphrite. La patiente disait avoir une éruption cutanée bilatérale des membres inférieurs et une douleur intermittente au genou avec enflure depuis trois mois, ainsi que des épisodes de fièvre subjective au cours des deux dernières années. L'analyze d'urine a révélé une hématurie microscopique persistante et un taux de créatinine élevé à 94 umol/L (59 umol/L initialement). Ses premiers examens ont montré une élévation de la CRP et du facteur rhumatoïde, ainsi qu'un titer d'ANA faiblement positif. Diagnostic: La biopsie rénale était compatible avec une glomérulonéphrite mésangiale proliférative à complexe immun. La microscopie optique a démontré une hypercellularité mésangiale diffuse globale. L'immunofluorescence des cellules mésangiales a révélé des traces d'IgG, de kappa, d'IgM, de chaînes lambda et de fragments C1q 1+ et de C3 2+. La microscopie électronique a montré des dépôts denses aux électrons en localization mésangiale. Ces résultats ont été jugés comme plus conformes à la néphrite lupique mésangiale proliférative; on a toutefois reconnu que des résultats cliniques et de laboratoire appuyant ce diagnostic faisaient défaut. Intervention: Après un traitement par prednisone orale, les symptômes de la patiente se sont résorbés et sa fonction rénale s'est améliorée. Il ne lui a toutefois pas été possible de cesser complètement la prednisone sans une réapparition de ses symptômes. D'autres traitements immunosuppresseurs ont été mis à l'essai, mais la patiente est restée dépendante des stéroïdes en raison de poussées de la maladie liées aux réductions de la dose de prednisone. Cette réponse atypique au traitement a mené l'équipe soignante à envisager d'autres diagnostics. Des examens supplémentaires ont révélé une sérologie positive pour la fièvre Q (IgG phase I 1:1892; IgG phase II 1:8192; IgM phase I et II <1:16). La patiente a reçu un diagnostic de fièvre Q chronique et a été traitée avec de la doxycycline et de l'hydroxychloroquine. Résultats: La patiente a reçu le traitement pendant deux ans au cours desquels ses symptômes se sont résorbés, son taux de créatinine est revenu à la valeur initiale et l'hématurie est disparue. Après l'arrêt du traitement, ses titres d'IgM de fièvre Q ont augmenté et la patiente a dû reprendre indéfiniment le traitement par doxycycline et hydroxychloroquine. Enseignements tirés: (1) Pour garantir un diagnostic précis et le traitement approprié, il est important d'envisager un diagnostic différentiel étendu en présence de caractéristiques cliniques atypiques ou d'une réponse inattendue au traitement. (2) L'amélioration clinique liée au traitement immunosuppresseur n'exclut pas une étiologie infectieuse de la glomérulonéphrite.
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Background and Objective: Lung-protective mechanical ventilation is known to attenuate ventilator-associated lung injury (VALI), but often at the expense of hypoventilation and hypercapnia. It remains unclear whether the main mechanism by which VALI is attenuated is a product of limiting mechanical forces to the lung during ventilation, or a direct biological effect of hypercapnia. Methods: Acute lung injury (ALI) was induced in 60 anesthetized rats by the instillation of 1.25 M HCl into the lungs via tracheostomy. Ten rats each were randomly assigned to one of six experimental groups and ventilated for 4 h with: 1) Conventional HighV E Normocapnia (high VT, high minute ventilation, normocapnia), 2) Conventional Normocapnia (high VT, normocapnia), 3) Protective Normocapnia (VT 8 ml/kg, high RR), 4) Conventional iCO 2 Hypercapnia (high VT, low RR, inhaled CO2), 5) Protective iCO 2 Hypercapnia (VT 8 ml/kg, high RR, added CO2), 6) Protective endogenous Hypercapnia (VT 8 ml/kg, low RR). Blood gasses, broncho-alveolar lavage fluid (BALF), and tissue specimens were collected and analyzed for histologic and biologic lung injury assessment. Results: Mild ALI was achieved in all groups characterized by a decreased mean PaO2/FiO2 ratio from 428 to 242 mmHg (p < 0.05), and an increased mean elastance from 2.46 to 4.32 cmH2O/L (p < 0.0001). There were no differences in gas exchange among groups. Wet-to-dry ratios and formation of hyaline membranes were significantly lower in low VT groups compared to conventional tidal volumes. Hypercapnia reduced diffuse alveolar damage and IL-6 levels in the BALF, which was also true when CO2 was added to conventional VT. In low VT groups, hypercapnia did not induce any further protective effect except increasing pulmonary IL-10 in the BALF. No differences in lung injury were observed when hypercapnia was induced by adding CO2 or decreasing minute ventilation, although permissive hypercapnia decreased the pH significantly and decreased liver histologic injury. Conclusion: Our findings suggest that low tidal volume ventilation likely attenuates VALI by limiting mechanical damage to the lung, while hypercapnia attenuates VALI by limiting pro-inflammatory and biochemical mechanisms of injury. When combined, both lung-protective ventilation and hypercapnia have the potential to exert an synergistic effect for the prevention of VALI.
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RATIONALE: Primary hyperoxaluria (PH) is a rare autosomal recessive disorder more commonly diagnosed in children or adolescents. Owing to its rarity and heterogeneous phenotype, it is often underrecognized, resulting in delayed diagnosis, including diagnosis after end-stage kidney disease (ESKD) has occurred or recurrence after kidney-only transplantation. CASE PRESENTATION: A 40-year-old Caucasian Canadian woman with a history of recurrent nephrolithiasis since age 19 presented with ESKD and cutaneous symptoms. She had no known prior kidney disease and no family history of kidney disease or nephrolithiasis. DIAGNOSIS: A diagnosis of primary hyperoxaluria type 1 (PH1) due to homozygous splice donor mutation (AGXT c.680+1G>A) was made with kidney and cutaneous pathology demonstrating calcium oxalate deposition and ultrasound suggestive of nephrocalcinosis. INTERVENTIONS: She was initiated on frequent, high-efficiency, high-flux conventional hemodialysis and oral pyridoxine. Lumasiran was added 11 months later, after she developed bilateral swan-neck deformities. OUTCOMES: After 14 months of high-intensity dialysis and 3 months of lumasiran, there have been no signs of renal recovery, and extra-renal involvement has increased with progressive swan-neck deformities, reduced cardiac systolic function, and pulmonary hypertension. The patient has been waitlisted for kidney-liver transplantation. TEACHING POINTS: This case report describes an adult presentation of PH1. The case highlights the importance of timely workup of metabolic causes of recurrent nephrolithiasis or nephrocalcinosis in adults which can be a presenting sign of PH and genetic testing for PH to facilitate early diagnosis and treatment especially in the era of novel therapeutics that may alter disease course and outcomes. The case also demonstrates the value of testing for PH in adults presenting with unexplained ESKD and a history of recurrent nephrolithiasis or nephrocalcinosis due to implications for organ transplantation strategy and presymptomatic family screening.
JUSTIFICATION: L'hyperoxalurie primaire (HP) est un trouble récessif autosomique rare plus souvent rencontré chez les enfants ou les adolescents. En raison de sa rareté et de son phénotype hétérogène, cette affection est fréquemment sous-reconnue, ce qui entraîne un retard dans le diagnostic, et ce, même après l'apparition d'une insuffisance rénale terminale (IRT) ou une récidive suivant une greffe simple de rein. PRÉSENTATION DU CAS: Nous présentons le cas d'une Canadienne de race blanche âgée de 40 ans avec des antécédents de néphrolithiase récurrente depuis l'âge de 19 ans. La patiente était atteinte d'IRT et présentait des symptômes cutanés. Elle n'avait aucun antécédent connu de maladie rénale ou antécédent familial de maladie rénale ou de néphrolithiase. DIAGNOSTIC: Une pathologie rénale et cutanée montrant des dépôts d'oxalate de calcium et une échographie suggérant une néphrocalcinose ont permis de poser un diagnostic d'hyperoxalurie primaire de type 1 (HP1) due à une mutation de donneur d'épissage homozygote (AGXT c.680+1G>A). INTERVENTIONS: La patiente a amorcé des traitements d'hémodialyse conventionnelle à grande fréquence, à haut rendement et à flux élevé, et a reçu de la pyridoxine par voie orale. Un traitement par lumasiran a été ajouté 11 mois plus tard, après le développement de déformations bilatérales en col de cygne. RÉSULTATS: Après quatorze mois de dialyze à haute intensité et trois mois de lumasiran, aucun signe de récupération rénale n'a été observé. L'intervention d'épuration extra-rénale a été augmentée en raison de déformations progressives en col de cygne, d'une réduction de la fonction cardiaque systolique et d'une hypertension pulmonaire. La patiente a été placée sur la liste d'attente pour une transplantation rénale et hépatique. ENSEIGNEMENTS TIRÉS: Ce rapport de cas décrit une présentation adulte d'HP1. Ce cas souligne l'importance de traiter rapidement les causes métaboliques de la néphrolithiase ou de la néphrocalcinose récidivante chez les adultes, car celles-ci peuvent être des signes d'hyperoxalurie primaire (HP). Ce cas souligne en outre l'importance de procéder à des tests génétiques pour l'HP afin de permettre le diagnostic et le traitement précoces, en particulier à l'ère de nouveaux traitements susceptibles d'infléchir l'évolution et les résultats de la maladie. Enfin, il démontre la valeur du dépistage de l'HP chez les adultes présentant une IRT inexpliquée et des antécédents de néphrolithiase ou de néphrocalcinose récidivante, en raison de ses implications sur la stratégie de transplantation d'organes et sur le dépistage pré-symptomatique de la famille.
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RATIONALE: Alemtuzumab is a monoclonal antibody approved for the treatment of relapsing-remitting multiple sclerosis (RRMS). Many autoimmune-mediated adverse events have been associated with alemtuzumab, including renal-limited anti-glomerular basement membrane (GBM) disease. PRESENTING CONCERN: A 52-year-old female with RRMS presented with acute kidney injury 39 months after receiving 1 cycle of alemtuzumab. She had a history of alemtuzumab-associated hypothyroidism and thrombocytopenia, urinary tract infections, and chronically abnormal urinalyses. DIAGNOSIS: A diagnosis of renal-limited anti-GBM disease was made based on renal biopsy and positive anti-GBM serology. Alemtuzumab was thought to be the trigger of the anti-GBM disease as there were no other exposures or serologic findings suggesting other causes. INTERVENTIONS: She was treated with corticosteroids, cyclophosphamide, and plasmapheresis. She required hemodialysis for acute renal failure. OUTCOMES: Despite treatment, the patient's renal function did not recover. She remained dialysis-dependent and anti-GBM antibody titers remained elevated 6 months after presentation. TEACHING POINTS: Anti-GBM disease is a life-altering adverse event that can be associated with alemtuzumab. Our case highlights the limitations of monitoring urinalyses as a trigger for anti-GBM antibody testing in patients who have received alemtuzumab and have baseline abnormal urinalyses; such patients may require further protocolized anti-GBM antibody testing, although the optimal frequency of such antibody screening remains unclear.
JUSTIFICATION: L'alemtuzumab est un anticorps monoclonal approuvé pour le traitement de la sclérose en plaque récurrente-rémittente (SPRR). De nombreux événements indésirables à médiation auto-immune ont été associés à ce traitement, notamment la glomérulonéphrite auto-immune (maladie anti-MBG). PRÉSENTATION DU CAS: Une femme de 52 ans atteinte de SPRR présentant une insuffisance rénale aiguë 39 mois après avoir reçu un cycle d'alemtuzumab. La patiente avait des antécédents d'hypothyroïdie et de thrombocytopénie liées à la prise d'alemtuzumab, en plus de présenter des tests urinaires anormaux et de souffrir d'infections des voies urinaires de façon chronique. DIAGNOSTIC: Un diagnostic de glomérulonéphrite auto-immune a été établi sur la base d'une biopsie rénale et d'une sérologie anti-MBG positive. On a suspecté l'alemtuzumab d'être à l'origine de la glomérulonéphrite auto-immune puisqu'aucun résultat sérologique ou exposition ne suggérait d'autres causes. INTERVENTIONS: La patiente a été traitée aux corticostéroïdes, au cyclophosphamide et par plasmaphérèse. L'insuffisance rénale aiguë a requis un traitement d'hémodialyse. RÉSULTATS: Malgré le traitement, la fonction rénale de la patiente ne s'est pas rétablie. La patiente a dû poursuivre les traitements de dialyse et ses titres d'anticorps demeuraient élevés six mois après la présentation des symptômes. LEÇONS TIRÉES: La glomérulonéphrite auto-immune est un événement indésirable aux conséquences dévastatrices et cette affection peut être associée à la prise d'alemtuzumab. Notre cas met en lumière les limites du suivi des analyses urinaires comme critère de dépistage des anticorps anti-MBG chez les patients ayant reçu de l'alemtuzumab et dont les analyses urinaires préalables sont anormales. Ces patients pourraient nécessiter des tests de détection des anticorps anti-MBG supplémentaires, bien que leur fréquence optimale demeure incertaine.
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BACKGROUND: Current uncertainties about the similarity between human diseases and their experimental models are hampering the development of new therapies. This is especially the case for diabetic kidney disease (DKD), the most common cause of end-stage kidney disease. To better understand the nature of the commonality between humans and their mouse models, we posed the question: in diabetic kidney disease are transcriptional profiles primarily disease-specific or species-specific? METHODS: We performed a meta-comparison of the glomerular transcriptomic characteristics of 133 human and 66 mouse samples including five human kidney diseases and five mouse models, validating expression patterns of a central node by immunohistochemistry. FINDINGS: Principal component analysis controlled for mouse background, revealed that gene expression changes in glomeruli from humans with DKD are more similar to those of diabetic mice than they are to other human glomerular diseases. This similarity enabled the construction of a discriminatory classifier that distinguishes diabetic glomeruli from other glomerular phenotypes regardless of their species of origin. To identify where the commonality between mice and humans with diabetes lies, networks of maximally perturbed protein interactions were examined, identifying a central role for the epidermal growth factor receptor (EGFR). By immunohistochemical staining, we found EGFR to be approximately doubled in its glomerular expression in both humans and mice. INTERPRETATION: These findings indicate that diabetic mouse models do mimic some of the features of human kidney disease, at least with respect to their glomerular transcriptomic signatures, and they identify EGFR as being a central player in this inter-species overlap.
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Nefropatías Diabéticas , Fallo Renal Crónico , Glomérulos Renales , Transcriptoma , Animales , Nefropatías Diabéticas/diagnóstico , Nefropatías Diabéticas/genética , Modelos Animales de Enfermedad , Receptores ErbB/metabolismo , Humanos , Fallo Renal Crónico/diagnóstico , Fallo Renal Crónico/genética , Glomérulos Renales/metabolismo , RatonesRESUMEN
Novel tools are needed to improve diagnostic accuracy and risk prediction in BK virus nephropathy (BKVN). We assessed the utility of intragraft gene expression testing for these purposes. Eight hundred genes were measured in 110 archival samples, including a discovery cohort of native kidney BKVN (n = 5) vs pure T cell-mediated rejection (TCMR; n = 10). Five polyomavirus genes and seven immune-related genes (five associated with BKVN and two associated with TCMR) were significantly differentially expressed between these entities (FDR < 0.05). These three sets of genes were further evaluated in samples representing a spectrum of BK infection (n = 25), followed by a multicenter validation cohort of allograft BKVN (n = 60) vs TCMR (n = 10). Polyomavirus 5-gene set expression reliably distinguished BKVN from TCMR (validation cohort AUC = 0.992), but the immune gene sets demonstrated suboptimal diagnostic performance (AUC ≤ 0.720). Within the validation cohort, no significant differences in index biopsy gene expression were identified between BKVN patients demonstrating resolution (n = 35), persistent infection (n = 14) or de novo rejection (n = 11) 6 months following a standardized reduction in immunosuppression. These results suggest that, while intragraft polyomavirus gene expression may be useful as an ancillary diagnostic for BKVN, assessment for concurrent TCMR and prediction of clinical outcome may not be feasible with current molecular tools.
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Virus BK , Enfermedades Renales , Trasplante de Riñón , Infecciones por Polyomavirus , Infecciones Tumorales por Virus , Virus BK/genética , Expresión Génica , Rechazo de Injerto/etiología , Rechazo de Injerto/genética , Humanos , Riñón , Enfermedades Renales/diagnóstico , Enfermedades Renales/genética , Trasplante de Riñón/efectos adversos , Infecciones por Polyomavirus/diagnóstico , Medición de Riesgo , Linfocitos T , Infecciones Tumorales por Virus/diagnósticoRESUMEN
Since 2014, programmed cell death protein 1 (PD-1)/programmed cell death ligand 1 (PD-L1) checkpoint inhibitors have been approved by various regulatory agencies for the treatment of multiple cancers including melanoma, lung cancer, urothelial carcinoma, renal cell carcinoma, head and neck cancer, classical Hodgkin lymphoma, colorectal cancer, gastroesophageal cancer, hepatocellular cancer, and other solid tumors. Of these approved drug/disease combinations, a subset also has regulatory agency-approved, commercially available companion/complementary diagnostic assays that were clinically validated using data from their corresponding clinical trials. The objective of this document is to provide evidence-based guidance to assist clinical laboratories in establishing fit-for-purpose PD-L1 biomarker assays that can accurately identify patients with specific tumor types who may respond to specific approved immuno-oncology therapies targeting the PD-1/PD-L1 checkpoint. These recommendations are issued as 38 Guideline Statements that address (i) assay development for surgical pathology and cytopathology specimens, (ii) reporting elements, and (iii) quality assurance (including validation/verification, internal quality assurance, and external quality assurance). The intent of this work is to provide recommendations that are relevant to any tumor type, are universally applicable and can be implemented by any clinical immunohistochemistry laboratory performing predictive PD-L1 immunohistochemistry testing.
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Antineoplásicos Inmunológicos/uso terapéutico , Antígeno B7-H1/metabolismo , Biomarcadores/metabolismo , Inmunoterapia/métodos , Neoplasias/terapia , Antígeno B7-H1/antagonistas & inhibidores , Canadá , Técnicas de Laboratorio Clínico , Medicina Basada en la Evidencia , Humanos , Inmunohistoquímica , Neoplasias/diagnóstico , Neoplasias/inmunología , Selección de Paciente , Guías de Práctica Clínica como Asunto , Valor Predictivo de las Pruebas , Pronóstico , Garantía de la Calidad de Atención de SaludRESUMEN
AIMS/HYPOTHESIS: Long non-coding RNAs (lncRNAs) are garnering increasing attention for their putative roles in the pathogenesis of chronic diseases, including diabetic kidney disease (DKD). However, much about in vivo lncRNA functionality in the adult organism remains unclear. To better understand lncRNA regulation and function in DKD, we explored the effects of the modular scaffold lncRNA HOTAIR (HOX antisense intergenic RNA), which approximates chromatin modifying complexes to their target sites on the genome. METHODS: Experiments were performed in human kidney tissue, in mice with streptozotocin-induced diabetes, the db/db mouse model of type 2 diabetes, podocyte-specific Hotair knockout mice and conditionally immortalised mouse podocytes. RESULTS: HOTAIR was observed to be expressed by several kidney cell-types, including glomerular podocytes, in both human and mouse kidneys. However, knockout of Hotair from podocytes had almost no effect on kidney structure, function or ultrastructure. Glomerular HOTAIR expression was found to be increased in human DKD, in the kidneys of mice with streptozotocin-induced diabetes and in the kidneys of db/db mice. Likewise, exposure of cultured mouse podocytes to high glucose caused upregulation of Hotair expression, which occurred in a p65-dependent manner. Although HOTAIR expression was upregulated in DKD and in high glucose-exposed podocytes, its knockout did not alter the development of kidney damage in diabetic mice. Rather, in a bioinformatic analysis of human kidney tissue, HOTAIR expression closely paralleled the expression of its genic neighbour, HOXC11, which is important to developmental patterning but which has an uncertain role in the adult kidney. CONCLUSIONS/INTERPRETATION: Many lncRNAs have been found to bind to the same chromatin modifying complexes. Thus, there is likely to exist sufficient redundancy in the system that the biological effects of dysregulated lncRNAs in kidney disease may often be inconsequential. The example of the archetypal scaffold lncRNA, HOTAIR, illustrates how lncRNA dysregulation may be a bystander in DKD without necessarily contributing to the pathogenesis of the condition. In the absence of in vivo validation, caution should be taken before ascribing major functional roles to single lncRNAs in the pathogenesis of chronic diseases.
Asunto(s)
Nefropatías Diabéticas/metabolismo , Regulación de la Expresión Génica , ARN Largo no Codificante/metabolismo , Animales , Tipificación del Cuerpo , Cromatina/metabolismo , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Proteínas de Homeodominio/metabolismo , Humanos , Hibridación in Situ , Glomérulos Renales/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Podocitos/citología , Podocitos/metabolismo , ARN Largo no Codificante/genéticaAsunto(s)
Tasa de Filtración Glomerular/fisiología , Riñón/fisiopatología , Nefritis Intersticial/diagnóstico , Oxalatos/toxicidad , Rheum/toxicidad , Anciano de 80 o más Años , Biopsia , Creatinina/orina , Humanos , Riñón/patología , Masculino , Nefritis Intersticial/etiología , Nefritis Intersticial/fisiopatología , Nefritis Intersticial/terapia , Oxalatos/análisis , Oxalatos/metabolismo , Diálisis Renal , Rheum/química , Factores de RiesgoRESUMEN
The posttranslational histone modifications that epigenetically affect gene transcription extend beyond conventionally studied methylation and acetylation patterns. By examining the means by which podocytes influence the glomerular endothelial phenotype, we identified a role for phosphorylation of histone H3 on serine residue 10 (phospho-histone H3Ser10) in mediating endothelial activation in diabetes. Culture media conditioned by podocytes exposed to high glucose caused glomerular endothelial vascular cell adhesion protein 1 (VCAM-1) upregulation and was enriched for the chemokine CCL2. A neutralizing anti-CCL2 antibody prevented VCAM-1 upregulation in cultured glomerular endothelial cells, and knockout of the CCL2 receptor CCR2 diminished glomerular VCAM-1 upregulation in diabetic mice. CCL2/CCR2 signaling induced glomerular endothelial VCAM-1 upregulation through a pathway regulated by p38 mitogen-activated protein kinase, mitogen- and stress-activated protein kinases 1/2 (MSK1/2), and phosphorylation of H3Ser10, whereas MSK1/2 inhibition decreased H3Ser10 phosphorylation at the VCAM1 promoter. Finally, increased phospho-histone H3Ser10 levels were observed in the kidneys of diabetic endothelial nitric oxide synthase knockout mice and in the glomeruli of humans with diabetic kidney disease. These findings demonstrate the influence that histone protein phosphorylation may have on gene activation in diabetic kidney disease. Histone protein phosphorylation should be borne in mind when considering epigenetic targets amenable to therapeutic manipulation in diabetes.
Asunto(s)
Nefropatías Diabéticas/metabolismo , Endotelio Vascular/metabolismo , Histonas/metabolismo , Transducción de Señal/fisiología , Animales , Células Endoteliales/metabolismo , Humanos , Glomérulos Renales/metabolismo , Ratones , Ratones Noqueados , Óxido Nítrico Sintasa de Tipo III/metabolismo , Fosforilación , Podocitos/metabolismo , Regiones Promotoras Genéticas , Receptores CCR2/genética , Receptores CCR2/metabolismo , Molécula 1 de Adhesión Celular Vascular/genética , Molécula 1 de Adhesión Celular Vascular/metabolismoRESUMEN
To contend with the deleterious effects of accumulating misfolded protein aggregates or damaged organelles cells rely on a system of quality control processes, among them the autophagy-lysosome pathway. This pathway is itself controlled by a master regulator transcription factor termed transcription factor EB (TFEB). When TFEB localizes to the cell nucleus it promotes the expression of a number of genes involved in protein clearance. Here, we set out to determine (1) whether TFEB expression is altered in chronic kidney disease (CKD); (2) whether inhibition of the cytosolic deacetylase histone deacetylase 6 (HDAC6) affects TFEB acetylation and nuclear localization; and (3) whether HDAC6 inhibition, in turn, alters the natural history of experimental CKD. TFEB mRNA and protein levels were observed to be diminished in the kidneys of humans with diabetic kidney disease, accompanied by accumulation of the protein aggregate adaptor protein p62 in tubule epithelial cells. In cultured NRK-52E cells, HDAC6 inhibition with the small molecule inhibitor Tubastatin A acetylated TFEB, increasing TFEB localization to the nucleus and attenuating cell death. In a rat model of CKD, Tubastatin A prevented the accumulation of misfolded protein aggregates in tubule epithelial cells, attenuated proteinuria progression, limited tubule cell death and diminished tubulointerstitial collagenous matrix deposition. These findings point to the common occurrence of dysregulated quality control processes in CKD and they suggest that TFEB downregulation may contribute to tubule injury in CKD. They also identify a regulatory relationship between HDAC6 and TFEB. HDAC6 inhibitors and TFEB activators both warrant further investigation as treatments for CKD.
RESUMEN
Histone protein modifications control fate determination during normal development and dedifferentiation during disease. Here, we set out to determine the extent to which dynamic changes to histones affect the differentiated phenotype of ordinarily quiescent adult glomerular podocytes. To do this, we examined the consequences of shifting the balance of the repressive histone H3 lysine 27 trimethylation (H3K27me3) mark in podocytes. Adriamycin nephrotoxicity and subtotal nephrectomy (SNx) studies indicated that deletion of the histone methylating enzyme EZH2 from podocytes decreased H3K27me3 levels and sensitized mice to glomerular disease. H3K27me3 was enriched at the promoter region of the Notch ligand Jag1 in podocytes, and derepression of Jag1 by EZH2 inhibition or knockdown facilitated podocyte dedifferentiation. Conversely, inhibition of the Jumonji C domain-containing demethylases Jmjd3 and UTX increased the H3K27me3 content of podocytes and attenuated glomerular disease in adriamycin nephrotoxicity, SNx, and diabetes. Podocytes in glomeruli from humans with focal segmental glomerulosclerosis or diabetic nephropathy exhibited diminished H3K27me3 and heightened UTX content. Analogous to human disease, inhibition of Jmjd3 and UTX abated nephropathy progression in mice with established glomerular injury and reduced H3K27me3 levels. Together, these findings indicate that ostensibly stable chromatin modifications can be dynamically regulated in quiescent cells and that epigenetic reprogramming can improve outcomes in glomerular disease by repressing the reactivation of developmental pathways.
Asunto(s)
Nefropatías Diabéticas/metabolismo , Histonas/metabolismo , Podocitos/metabolismo , Animales , Nefropatías Diabéticas/patología , Proteína Potenciadora del Homólogo Zeste 2/metabolismo , Femenino , Histona Demetilasas/metabolismo , Humanos , Proteína Jagged-1/metabolismo , Histona Demetilasas con Dominio de Jumonji/metabolismo , Masculino , Metilación , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , Proteínas Nucleares/metabolismo , Podocitos/patologíaRESUMEN
BACKGROUND: An adequate renal biopsy is essential for diagnosis and treatment of medical renal disease. OBJECTIVE: We evaluated two initiatives to improve adequacy of renal biopsy samples at our centre. DESIGN: Retrospective determination of renal biopsy adequacy. SETTING: Queen Elizabeth II Health Sciences Centre. PATIENTS: Patients undergoing medical renal biopsies. MEASUREMENTS: Renal biopsy adequacy. METHODS: The first initiative was to restrict the performance of biopsies to a smaller group of radiologists and to include a comment on biopsy adequacy in every pathology report. The second initiative was to introduce on-site adequacy assessment by a medical laboratory technologist. Native renal and allograft biopsy adequacies were calculated for three periods: 1) baseline, October 2005 to September 2006; 2) after implementation of the first initiative, January 2007 to September 2011; and 3) after implementation of the second initiative, October 2011 to September 2012. A subset of native renal biopsies was examined to determine if there was a relationship between adequacy and number of passes. RESULTS: The percentages of adequate native renal biopsies during the first, second, and third periods were 31%, 72% and 90%, respectively. This represents a significant increase (40%, p < 0.0001) in adequacy following the first initiative, and another significant increase (18%, p = 0.0003) following the second initiative. The percentages of adequate renal allograft biopsies during the first, second, and third periods were 75%, 56% and 69%, respectively. These changes in adequacy were not statistically significant. In the subset of native renal biopsies examined, a biopsy comprising more than three cores was not associated with increase in adequacy. LIMITATIONS: The most important limitation is the lack of generally accepted and applied adequacy criteria limiting generalizability of our findings. CONCLUSIONS: Restricting the performance of biopsies to subspecialist operators, including an adequacy statement in the renal biopsy report and on-site adequacy assessment were effective in significantly improving native renal biopsy adequacy. This improvement appeared unrelated to an increase in the number of passes taken with a biopsy needle. Neither initiative improved the low adequacy of allograft biopsies.
CONTEXTE: L'importance que joue la biopsie percutanée dans le diagnostic et le traitement des maladies rénales est un fait bien connu. Un spécimen adéquat est nécessaire pour une interprétation correcte des résultats. La surveillance de la qualité de biopsie reste une pratique importante de contrôle de la qualité. OBJECTIF DE L'ÉTUDE: Deux initiatives visant une meilleure qualité des échantillons rénaux pour la biopsie ont été évaluées à notre centre. TYPE D'ÉTUDE: Détermination rétrospective de la qualité de la biopsie rénale. CONTEXTE: Le Queen Elizabeth II Health Sciences Centre. PATIENTS: Patients subissant des biopsies rénales. MESURES: Qualité de la biopsie rénale. MÉTHODES: La première initiative visait à restreindre la réalisation des biopsies à un petit groupe de radiologues et d'ajouter un commentaire sur la qualité de la biopsie dans chaque rapport de pathologie. La deuxième initiative visait à implanter l'évaluation de cette qualité sur place, par l'intermédiaire d'un technicien de laboratoire médical. La qualité des biopsies de reins natifs et d'allogreffes ont été calculées en trois temps : 1) mesure de référence: d'octobre 2005 à septembre 2006; 2) après l'implantation de la première initiative, de janvier 2007 à septembre 2011; 3) après l'implémentation de la deuxième initiative, d'octobre 2011 à septembre 2012. Un sous-ensemble de biopsies effectuées sur des reins natifs a été examiné afin de déterminer s'il existait un lien entre la qualité et le nombre de ponctions. RÉSULTATS: Les pourcentages d'une qualité adéquate des biopsies de reins natifs des trois phases étaient de 31%, 72% et 90%, respectivement. On remarque des hausses de la qualité des biopsies importantes après la première initiative (40%, p < 0,0001) et après la deuxième (18%, p = 0,0003). Les pourcentages d'une qualité adéquate des biopsies d'allogreffes rénales des trois étapes étaient de 75%, 56% et 69%, respectivement; ces changements n'étaient pas statistiquement significatifs. Dans le sous-ensemble des biopsies de reins natifs, celles de plus de trois échantillons n'étaient pas associées avec une hausse de la qualité. LIMITES DE L'ÉTUDE: Parmi les limites de l'étude, on compte les données lacunaires concernant : les complications postopératoires; l'analyse détaillée des caractéristiques des radiologistes avant et après la première intervention; certaines données permettant de mesurer l'effet de la deuxième intervention indépendamment de son lien avec la première intervention; un consensus sur les critères de la qualité des biopsies de reins natifs et d'allogreffes rénales à des fins de comparaisons entre les différentes institutions, et qui s'est répercuté sur la généralisabilité de nos résultats (provenant d'un seul centre) à d'autres établissements. Le manque de puissance statistique a limité la possibilité de détecter certaines différences entre les sous-groupes. CONCLUSIONS: La restriction de la réalisation des biopsies par une sous-spécialité, l'ajout d'un commentaire sur la qualité de la biopsie rénale dans le rapport, et l'évaluation sur place de la qualité ont eu l'effet d'améliorer de façon significative la proportion de biopsies rénales natives avec une qualité adéquate. Cette amélioration ne semble pas liée à une hausse du nombre de ponctions faites avec une aiguille à biopsie. Aucune des initiatives n'a amélioré la basse qualité des biopsies d'allogreffes.
RESUMEN
OBJECTIVES: Formalin-fixed, paraffin-embedded unstained archived diagnostic tissue sections are frequently exchanged between clinical laboratories for immunohistochemical staining. The manner in which such sections are prepared represents a type of preanalytical variable that must be taken into account given the growing importance of immunohistochemical assays, especially predictive and prognostic tests, in personalized medicine. METHODS: Recommendations were derived from review of the literature and expert consensus of the Canadian Association of Pathologists-Association canadienne des pathologists National Standards Committee for High Complexity Testing/Immunohistochemistry. RESULTS: Relevant considerations include the type of glass slide on which to mount the unstained sections; the thickness of the tissue sections; the time from slide preparation to testing; the environment, particularly the temperature at which the unstained sections will be maintained prior to testing; the inclusion of on-slide positive control tissue where possible; and whether patient identifier(s) should be included on slide labels. CONCLUSIONS: Clear communication between requesting and releasing laboratories will facilitate the proper preparation of unstained sections and also ensure that applicable privacy considerations are addressed.