Michael D. Hughson

Nephron Number, Hypertension, Renal Disease, and Renal Failure

Essential hypertension is one of the most common diseases in the Western world, affecting about 26.4% of the adult population, and it is increasing (1). Its causes are heterogeneous and include genetic and environmental factors (2), but several observations point to an important role of the kidney in its genesis (3). In addition to variations in tubular transport mechanisms that could, for example, affect salt handling, structural characteristics of the kidney might also contribute to hypertension. The burden of chronic kidney disease is also increasing worldwide, due to population growth, increasing longevity, and changing risk factors. Although single-cause models of disease are still widely promoted, multideterminant or multihit models that can accommodate multiple risk factors in an individual or in a population are probably more applicable (4,5). In such a framework, nephron endowment is one potential determinant of disease susceptibility. Some time ago, Brenner and colleagues (6,7) proposed that lower nephron numbers predispose both to essential hypertension and to renal disease. They also proposed that hypertension and progressive renal insufficiency might be initiated and accelerated by glomerular hypertrophy and intraglomerular hypertension that develops as nephron number is reduced (8). In this review, we summarize data from recent studies that shed more light on these hypotheses. The data supply a new twist to possible mechanisms of the Barker hypothesis, which proposes that intrauterine growth retardation predisposes to chronic disease in later life (9). The review describes how nephron number is estimated and its range and some determinants and morphologic correlates. It then considers possible causes of low nephron numbers. Finally, associations of hypertension and renal disease with reduced nephron numbers are considered, and some potential clinical implications are discussed.

Nephron number, glomerular volume, renal disease and hypertension

Purpose of reviewTo discuss studies evaluating associations of glomerular number (Nglom) and glomerular volume with hypertension and kidney disease. Important findingsThe association of low Nglom with hypertension and renal insufficiency was described in the 1930s. Many investigators have noted loss of glomeruli with age, with most disappearing entirely, and have proposed that hypertension follows. In a recent German study, hypertensive patients had fewer glomeruli and larger mean glomerular volumes than nonhypertensive people. Among the 10-fold range of Nglom in our multiracial autopsy series, the lowest were in Australian Aborigines, who have the highest rates of renal failure. Nglom fell with age. There was a five-fold range in mean glomerular volume and considerable heterogeneity in individual glomerular volumes within a patient. Larger mean glomerular volume and greater individual glomerular volume heterogeneity correlated with lower Nglom, larger body size, hypertension, and black race. Hypertension increased with age and was marked by glomerular enlargement, intimal thickening and higher rates of glomerulosclerosis. In whites and Aborigines, but not in US blacks, lower Nglom was associated with hypertension, while robust numbers were highly protective. SummaryHigher mean glomerular volume and individual glomerular volume heterogeneity mark glomerular stress. Low Nglom is an important determinant of hypertension and renal disease. Many ‘missing’ nephrons have probably been lost during life, leaving little trace. Additional factors contribute to high rates of hypertension in blacks.

Pulmonary thrombotic arteriopathy in patients with sickle cell disease.

BACKGROUND Shortened life expectancy due to pulmonary hypertension (PH) is seen in 5% to 10% of patients with sickle cell disease. The principal factors suspected of causing PH are pulmonary thromboemboli (PE) and in situ arterial thrombosis. OBJECTIVE To investigate the possible role that PE or in situ arterial thrombosis play in the development of PH in sickle cell disease. METHODS Autopsies of 12 patients with sickle cell disease were correlated with clinical data from medical records. RESULTS Right ventricular hypertrophy was present in 9 of 12 patients. Six patients with right ventricular hypertrophy had thrombi in large elastic pulmonary arteries. All patients with elastic artery thrombi had fresh or organized thrombi in small muscular pulmonary arteries. Hypertensive small arterial changes were present in 5 of these 6 patients. Six patients showed no thrombi in elastic arteries. Among these 6 patients, 3 had right ventricular hypertrophy and recent and organized thrombi, as well as hypertensive changes in small arteries. One of these 3 patients demonstrated plexiform-like lesions and fibrinoid necrosis of small arteries. Three patients without right ventricular hypertrophy had pneumonia or pulmonary edema with no identifiable pulmonary artery pathology. CONCLUSIONS Arterial thrombosis with PH and cor pulmonale was regarded as the cause of death among most of these patients. Elastic artery thrombi are pulmonary thromboemboli, but pulmonary thromboemboli are always associated with widespread thrombosis of small arteries. Widespread thrombosis of small arteries alone was associated with PH in some cases. This finding suggests that pulmonary thromboemboli may be a late complication of PH and cor pulmonale and that an in situ thrombotic arteriopathy underlies the development of PH in most patients with sickle cell disease.

Antioxidant Treatment Prevents Renal Damage and Dysfunction and Reduces Arterial Pressure in Salt-Sensitive Hypertension

The goal of this study was to test the hypothesis that oxidative stress in Dahl salt-sensitive (SS) rats on a high-sodium intake contributes to the progression of renal damage, the decreases in renal hemodynamics, and the development of hypertension. We specifically studied whether antioxidant therapy, using vitamins C and E, could help prevent renal damage and glomerular filtration rate (GFR) and renal plasma flow reductions and attenuate the increases in arterial pressure. Thirty-three 7- to 8-week old Dahl SS/Rapp strain rats were placed on either a high-sodium (8%) or a low-sodium (0.3%) diet with or without vitamin E (111 IU/d) in the food and 98 mg/d vitamin C in the drinking water for 5 weeks. Rats were equipped with indwelling arterial and venous catheters at day 21. By day 35 in the rats with high-sodium diet, vitamin C and E treatment significantly decreased renal cortical and medullary &OV0151; release, mean arterial pressure, urinary protein excretion, glomerular necrosis, and renal tubulointerstitial damage. At this time, GFR significantly decreased in the high-sodium diet group (1.6±0.2 mL/min) when compared with either the high-sodium plus vitamins C and E (2.9±0.2 mL/min) or the low-sodium diet group (2.9±0.3 mL/min). In SS rats on high-sodium diet, renal plasma flow decreased 40%, and this reduced flow was restored by vitamin treatment. In Dahl salt-sensitive hypertension, increased oxidative stress plays an important role in the renal damage, decreases in renal hemodynamics, and increases in arterial pressure that occur. Antioxidant treatment with vitamins C and E improves renal dysfunction, lessens renal injury, and decreases arterial pressure in Dahl salt-sensitive hypertension.

A Common RET Variant Is Associated with Reduced Newborn Kidney Size and Function

Congenital nephron number varies five-fold among normal humans, and individuals at the lower end of this range may have an increased lifetime risk for essential hypertension or renal insufficiency; however, the mechanisms that determine nephron number are unknown. This study tested the hypothesis that common hypomorphic variants of the RET gene, which encodes a tyrosine kinase receptor critical for renal branching morphogenesis, might account for subtle renal hypoplasia in some normal newborns. A common single-nucleotide polymorphism (rs1800860 G/A) was identified within an exonic splicing enhancer in exon 7. The adenosine variant at mRNA position 1476 reduced affinity for spliceosome proteins, enhanced the likelihood of aberrant mRNA splicing, and diminished the level of functional transcript in human cells. In vivo, normal white newborns with an rs1800860(1476A) allele had kidney volumes 10% smaller and cord blood cystatin C levels 9% higher than those with the rs1800860(1476G) allele. These findings suggest that the RET(1476A) allele, in combination with other common polymorphic developmental genes, may account for subtle renal hypoplasia in a significant proportion of the white population. Whether this gene variant affects clinical outcomes requires further study.

The predictive value of p16INK4a and Hybrid Capture 2 human papillomavirus testing for high-grade cervical intraepithelial neoplasia

We performed p16 INK4a immunocytochemical analysis and Hybrid Capture 2 (HC2, Digene, Gaithersburg, MD) high-risk HPV testing on 210 abnormal SurePath (TriPath Imaging, Burlington, NC) Papanicolaou specimens diagnosed as low-grade squamous intraepithelial lesion (LSIL) or high-grade squamous intraepithelial lesion (HSIL). The results were compared with 121 follow-up biopsy specimens. p16 INK4a was positive in 57.9% of women with LSIL compared with 97.1% of women with HSIL. In contrast, HC2 testing was positive in 85.0% of women with LSIL and 86.4% of women with HSIL. The differences in the positive rates for16 INK4a between LSIL and HSIL was significant (P < .001), whereas for HC2 it was not (P = .264). In patients who had cervical biopsies following a cytologic diagnosis of LSIL, the positive predictive value (PPV) of p16 INK4a for a biopsy of cervical intraepithelial neoplasia grade 2 or 3 (CIN2/3; 33%) was significantly higher than the PPV of HC2 results (21%) (P < .001). Using liquid-based cytology specimens, p16 INK4a immunocytochemical analysis has a higher PPV than reflex HC2 HPV testing for identifying CIN2/3 among patients with LSIL and might be useful for selecting patients with LSIL for

Associations of Glomerular Number and Birth Weight With Clinicopathological Features of African Americans and Whites

BACKGROUND Hypertension and its cardiovascular complications affect African Americans more severely than whites, a disparity variously ascribed to low birth weight, low glomerular number, an exaggerated arteriolonephrosclerotic blood pressure response, and inflammation-induced oxidative stress. STUDY DESIGN Case series. SETTING AND PARTICIPANTS Autopsy kidneys of 107 African Americans and 87 whites aged 18 to 65 years at a single medical center between 1998 and 2005. Excluded were persons with known premorbid kidney disease; pathological findings of severe arterioarteriolonephrosclerosis, nodular and diffuse diabetic glomerulosclerosis, or nonischemic cardiomyopathy. PREDICTORS & OUTCOMES Associations of: (1) race, age, sex, birth weight, obesity, and glomerular number (predictors) with hypertension and death from coronary artery (CAD) and cerebrovascular disease (CVD; outcomes); and (2) age, blood pressure, and race (predictors) with arteriolonephrosclerotic changes, including chronic tubulointerstitial inflammation (outcomes). MEASUREMENTS Hypertension ascertained from chart review and heart weight. Cause of death determined from chart review and autopsy findings. Birth weight obtained from birth records (115 persons). Total glomerular number (N(glom)) estimated by using the dissector/fractionator technique. Arteriolosclerosis, glomerulosclerosis, cortical fibrosis, and chronic inflammation by using CD68 density were measured morphometrically. RESULTS 59 African Americans (55%) and 32 whites (37%) were classified as hypertensive. CAD and CVD were the cause of death in 64 (33%) and 18 persons (9%), respectively. By using multiple linear regression, birth weight (P < 0.001) and sex (P < 0.01), but not race (P = 0.3) or age (P = 0.2), predicted N(glom) (P < 0.001; adjusted r(2) = 0.176). Hypertension was associated with African American race (P = 0.04), older age (P < 0.001), and male sex (P = 0.01), but not with N(glom) (P = 0.9), body mass index (P = 0.9), or birth weight (P = 0.4). Hypertension was the only significant factor associated with CAD and CVD (P < 0.001 for both). Interactions of age and blood pressure with race showed that although African Americans had more severe hypertension (P < 0.001) and arteriolosclerosis (P = 0.01) at a younger age than whites, there were no significant racial differences in degrees of arteriolosclerosis, glomerulosclerosis, cortical fibrosis, or CD68 density for any level of increased blood pressure. LIMITATIONS The study is observational and descriptive. CONCLUSIONS The more severe hypertension found in African Americans could not be attributed to racial differences in N(glom) or birth weight. CAD and CVD death and increased arteriolonephrosclerosis, including CD68 density, were determined by using blood pressure without a significant interacting contribution from race.

Determinants of Glomerular Volume in Different Cortical Zones of the Human Kidney

Enlarged glomerular size is a feature of focal segmental glomerulosclerosis, obesity-related glomerulopathy, diabetic nephropathy, and hypertension. The distribution of glomerular volumes within different cortical zones and glomerular volume alterations with age and obesity may contribute to understanding the evolution of these diseases. We analyzed the distributions of volumes of individual glomeruli in the superficial, middle, and juxtamedullary cortex of normal human kidneys using the disector/Cavalieri method. Volumes (V(glom)) of 720 nonsclerotic glomeruli (30 per kidney, 10 per zone) were estimated in autopsy kidneys of 24 American men, 12 aged 20 to 30 yr and 12 aged 51 to 69 yr. Black and white individuals were represented equally. The range of individual V(glom) within subjects varied from two- to eight-fold. There were no significant zonal differences in V(glom) in the young or those with body surface area (BSA) < or = 2.11 m(2). In contrast, superficial glomeruli in the older age group, in those with BSA > 2.11 m(2), and in white subjects were significantly larger than juxtamedullary glomeruli. Black subjects tended toward larger V(glom) than white subjects, and this difference was significant and most marked in the juxtamedullary zone and independent of age, BSA, and glomerular number. There is a wide range in individual V(glom) in adults. BSA, race, and age independently influence V(glom) in different zones of the renal cortex. These findings might reflect processes of aging and susceptibility factors to renal disease.

Glomerular number and size variability and risk for kidney disease.

Purpose of reviewThis review discusses current understandings of variability in glomerular number and size, and the implications for renal health. Recent findingsThe quantitative microanatomy of the normal human kidney varies widely. Of greatest significance, total nephron number varies at least 13-fold, and several genes and environmental factors that regulate human nephron endowment have been identified. Full or partial deletion of more than 25 genes in mice has been shown to result in renal hypoplasia and, when measured, reduced nephron endowment. Many more will likely be identified. As would be expected, some gene abnormalities increase nephron endowment above that found in control mice. Glomerular volume also varies widely, both between and within kidneys, and increased heterogeneity of glomerular volume within kidneys is associated with risk factors for kidney disease, including birth weight, age, race, body size and hypertension. SummaryData from several human populations indicate that the quantitative microanatomy of the human kidney varies considerably: total glomerular number varies at least 13-fold, mean glomerular volume varies up to seven-fold and the volumes of individual glomeruli within single kidneys can vary as much as eight-fold. Human glomerular number, size and size distribution are being found to correlate with risk factors for kidney disease. The genetic and fetal environmental regulators of nephrogenesis, and thereby nephron endowment, are being rapidly identified and will provide the bases for future clinical interventions. In contrast, the molecular regulation of glomerular size remains unclear.

Human papillomavirus genotyping and p16INK4a expression in cervical intraepithelial neoplasia of adolescents

Adolescents have high rates of human papillomavirus (HPV) infection, and persistent high-risk HPV infection can lead to the development of cervical cancer. The cyclin-dependent kinase inhibitor, p16INK4a is overexpressed in cervical intraepithelial neoplasia (CIN), probably due to a persistent and integrated HPV infection. This study investigated p16INK4a expression, grades of CIN, and high-risk HPV infection in adolescent cervical biopsies. Biopsies were immunohistochemically stained for p16INK4a. The presence of wide-spectrum, low-risk, or high-risk HPV was determined by amplifying DNA extracted from the cervical biopsies. Biopsies were classified as cervicitis, 15 cases; CIN 1, 48 cases; CIN 2, 46 cases, and CIN 3, 52 cases. The distribution of p16INK4a staining was graded as patchy, diffuse basal, and diffuse full thickness. Pearsons χ2 tests analyzed the relationships between p16INK4a staining, HPV infection, and CIN. Biopsies of cervicitis were negative for HPV and for p16INK4a expression. High-risk HPV 16, 18, and 31 increased from 18% in CIN 1 to 66% in CIN 2/3 (P<0.001). In CIN 1, p16INK4a was positive in 44% of biopsies with 35% showing patchy, 7% diffuse basal, and one case (2%) showing diffuse full thickness staining. In CIN 2/3, p16INK4a was positive in 97% of biopsies with 23% showing patchy, 21% diffuse basal, and 53% diffuse full thickness staining. The difference in the proportions of biopsies showing patchy p16INK4a staining in CIN 1 and diffuse full thickness staining in CIN 2/3 was significant (P<0.001). In CIN 1, 61% of high-risk HPV-positive biopsies were p16INK4a negative, while all high-risk HPV-positive CIN 2/3 biopsies were p16INK4a positive. Diffuse, full thickness p16INK4a expression discriminated low-grade from high-grade CIN and appears to be a marker of persistent high-risk HPV infection.