Role of fibulin-5 insufficiency and prolapse progression on murine vaginal biomechanical function.

The vagina plays a critical role in supporting the pelvic organs and loss of support leads to pelvic organ prolapse. It is unknown what microstructural changes influence prolapse progression nor how decreased elastic fibers contributes to vaginal remodeling and smooth muscle contractility. The objective for this study was to evaluate the effect of fibulin-5 haploinsufficiency, and deficiency with progressive prolapse on the biaxial contractile and biomechanical function of the murine vagina. Vaginas from wildtype (n = 13), haploinsufficient (n = 13), and deficient mice with grade 1 (n = 9) and grade 2 or 3 (n = 9) prolapse were explanted for biaxial contractile and biomechanical testing. Multiaxial histology (n = 3/group) evaluated elastic and collagen fiber microstructure. Western blotting quantified protein expression (n = 6/group). A one-way ANOVA or Kruskal-Wallis test evaluated statistical significance. Pearson's or Spearman's test determined correlations with prolapse grade. Axial contractility decreased with fibulin-5 deficiency and POP (p < 0.001), negatively correlated with prolapse grade (ρ = - 0.80; p < 0.001), and positively correlated with muscularis elastin area fraction (ρ = - 0.78; p = 0.004). Circumferential (ρ = 0.71; p < 0.001) and axial (ρ = 0.69; p < 0.001) vaginal wall stresses positively correlated with prolapse grade. These findings demonstrated that fibulin-5 deficiency and prolapse progression decreased vaginal contractility and increased vaginal wall stress. Future work is needed to better understand the processes that contribute to prolapse progression in order to guide diagnostic, preventative, and treatment strategies.

Single-cell transcriptome profiling of the vaginal wall in women with severe anterior vaginal prolapse.

Anterior vaginal prolapse (AVP) is the most common form of pelvic organ prolapse (POP) and has deleterious effects on women's health. Despite recent advances in AVP diagnosis and treatment, a cell atlas of the vaginal wall in AVP has not been constructed. Here, we employ single-cell RNA-seq to construct a transcriptomic atlas of 81,026 individual cells in the vaginal wall from AVP and control samples and identify 11 cell types. We reveal aberrant gene expression in diverse cell types in AVP. Extracellular matrix (ECM) dysregulation and immune reactions involvement are identified in both non-immune and immune cell types. In addition, we find that several transcription factors associated with ECM and immune regulation are activated in AVP. Furthermore, we reveal dysregulated cell-cell communication patterns in AVP. Taken together, this work provides a valuable resource for deciphering the cellular heterogeneity and the molecular mechanisms underlying severe AVP.

Polymorphism on chromosome 20p13 near the IDH3B gene is associated with uterine prolapse.

OBJECTIVES: Previous studies have found evidence for a genetic basis for pelvic organ prolapse (POP), but no genetic studies have differentiated between types of POP. This study investigated whether genetic variants in six previously suggested candidate loci for POP modify the risk of uterine prolapse (UP). STUDY DESIGN: One hundred patients, aged 30-55 years, who had undergone surgery due to total UP and 105 healthy controls were included in this study. After extracting the genomic DNA from peripheral blood, six single nucleotide polymorphisms (SNPs) previously identified for POP were genotyped, and association analysis was performed for contributing risk factors. RNA expression was determined from sacrouterine ligaments of patients and controls using quantitative reverse transcription polymerase chain reaction. RESULTS: The dominant genotype model for the T allele for SNP rs6051098 at the chromosome 20p13 locus was significant, and this remained significant with the risk factor regression model (p=0.046; odds ratio 1.93, 95 % confidence interval 1.01-3.66). The isocitrate dehydrogenase 3 beta (IDH3B) gene was the only potential candidate gene in the 20p13 locus that was significantly upregulated in sacrouterine biopsies in women with UP compared with controls (p = 0.034). CONCLUSION: To the best of the authors' knowledge, this is the first study to show that genetic risk factors contribute to UP, and suggested rs6051098 as the best candidate risk factor associated with UP. According to expression data in sacrouterine tissue, this study suggests that the IDH3B gene plays a role in the pathogenesis of UP.

Genome-wide association identifies seven loci for pelvic organ prolapse in Iceland and the UK Biobank.

Pelvic organ prolapse (POP) is a downward descent of one or more of the pelvic organs, resulting in a protrusion of the vaginal wall and/or uterus. We performed a genome-wide association study of POP using data from Iceland and the UK Biobank, a total of 15,010 cases with hospital-based diagnosis code and 340,734 female controls, and found eight sequence variants at seven loci associating with POP (P < 5 × 10-8); seven common (minor allele frequency >5%) and one with minor allele frequency of 4.87%. Some of the variants associating with POP also associated with traits of similar pathophysiology. Of these, rs3820282, which may alter the estrogen-based regulation of WNT4, also associates with leiomyoma of uterus, gestational duration and endometriosis. Rs3791675 at EFEMP1, a gene involved in connective tissue homeostasis, also associates with hernias and carpal tunnel syndrome. Our results highlight the role of connective tissue metabolism and estrogen exposure in the etiology of POP.

Expressions of homeobox, collagen and estrogen genes in women with uterine prolapse.

OBJECTIVE: Genetic contribution is thought to be involved in the pathophysiology of pelvic organ prolapse (POP). We aimed to study the gene expression profiles of the genes HomeoboxA11 (HOXA11), HomeoboxA13 (HOXA13), Collagen Type I (COL1A), Collagen Type III (COL3A), estrogen receptor genes (ESR1 and ESR2) of round (RL) and uterosacral ligaments (USL) in postmenopausal women with uterine prolapse. STUDY DESIGN: Gene expressions of 32 postmenopausal women with prolapse were analysed according to gene expressions of 8 postmenopausal women without prolapse. Quantitative real-time PCR (qRT-PCR) method was used for the detection of expression levels of the genes. Student's t-Test and Mann-Whitney U test were used for statistical analysis. RESULTS: In the USL specimens of all women with uterine prolapse HOXA13 and ESR1 gene expressions were decreased compared to controls (0.5 fold, p = 0.04 and 0.82 fold, p = 0.04, respectively). In the RL specimens, ESR2 gene expression was decreased 0.7 fold in women with prolapse when compared to controls (p = 0.04). In the USL specimens of women with advanced stages of prolapse (stage ≥3), HOXA13 and COL3A gene expressions were decreased compared to controls (0.44 fold, p = 0.043 and 0.39 fold, p = 0.045, respectively). In the RL specimens, ESR2 gene expression was decreased 0.65 fold in women with prolapse when compared to controls (p = 0.052). CONCLUSION: The significant decrease in the expression of the genes HOXA13, COL3A in the USL and ESR2 in the RL especially in advanced stages of prolapse, implicate that these gene expressions may play a role in the development of uterine prolapse.

Admixture mapping of pelvic organ prolapse in African Americans from the Women's Health Initiative Hormone Therapy trial.

Evidence suggests European American (EA) women have two- to five-fold increased odds of having pelvic organ prolapse (POP) when compared with African American (AA) women. However, the role of genetic ancestry in relation to POP risk is not clear. Here we evaluate the association between genetic ancestry and POP in AA women from the Women's Health Initiative Hormone Therapy trial. Women with grade 1 or higher classification, and grade 2 or higher classification for uterine prolapse, cystocele or rectocele at baseline or during follow-up were considered to have any POP (N = 805) and moderate/severe POP (N = 156), respectively. Women with at least two pelvic exams with no indication for POP served as controls (N = 344). We performed case-only, and case-control admixture-mapping analyses using multiple logistic regression while adjusting for age, BMI, parity and global ancestry. We evaluated the association between global ancestry and POP using multiple logistic regression. European ancestry at the individual level was not associated with POP risk. Case-only and case-control local ancestry analyses identified two ancestry-specific loci that may be associated with POP. One locus (Chromosome 15q26.2) achieved empirically-estimated statistical significance and was associated with decreased POP odds (considering grade ≥2 POP) with each unit increase in European ancestry (OR: 0.35; 95% CI: 0.30, 0.57; p-value = 1.48x10-5). This region includes RGMA, a potent regulator of the BMP family of genes. The second locus (Chromosome 1q42.1-q42.3) was associated with increased POP odds with each unit increase in European ancestry (Odds ratio [OR]: 1.69; 95% confidence interval [CI]: 1.28, 2.22; p-value = 1.93x10-4). Although this region did not reach statistical significance after considering multiple comparisons, it includes potentially relevant genes including TBCE, and ACTA1. Unique non-overlapping European and African ancestry-specific susceptibility loci may be associated with increased POP risk.

Microarray gene expression analysis of uterosacral ligaments in uterine prolapse.

OBJECTIVES: Pelvic organ prolapse (POP) is a major health problem that impairs the quality of life with a wide clinical spectrum. Since the uterosacral ligaments provide primary support for the uterus and the upper vagina, we hypothesize that the disruption of these ligaments may lead to a loss of support and eventually contribute to POP. DESIGN AND METHODS: In this study, we therefore investigated whether there are any differences in the transcription profile of uterosacral ligaments in patients with POP when compared to those of the control samples. Seventeen women with POP and 8 non-POP controls undergoing hysterectomy for benign conditions were included in the study. Affymetrix® Gene Chip microarrays (Human Hu 133 plus 2.0) were used for whole genome gene expression profiling analysis. RESULTS: There was 1 significantly down-regulated gene, NKX2-3 in patients with POP compared to the controls (p=4.28464e-013). KIF11 gene was found to be significantly down-regulated in patients with ≥3 deliveries compared to patients with <3 deliveries (p=0.0156237). UGT1A1 (p=2.43388e-005), SCARB1 (p=1.19001e-006) and NKX2-3 (p=2.17966e-013) genes were found to be significantly down-regulated in the premenopausal patients compared to the premenopausal controls. UGT1A1 gene was also found to be significantly down-regulated in the post menopausal patients compared to the postmenopausal controls (p=0.0005). CONCLUSION: This study provides evidence for a significant down-regulation of the genes that take role in cell cycle, proliferation and embryonic development along with cell adhesion process on the development of POP for the first time.

Genetic Mutation that May Contribute to Failure of Prolapse Surgery in White Women: A Case-Control Study.

OBJECTIVE: To identify a potential genetic basis for early failure after prolapse surgery. DESIGN: Case-control study (Canadian Task Force classification II). SETTING: This study was carried out in 1 academic community medical center referral practice, and all patients had surgery at 1 of 2 hospitals. PATIENTS: Ten women with early, multicompartment prolapse recurrence after robotic sacrocolpopexy compared with 40 control subjects with known success after the same procedure. INTERVENTIONS: Patients were treated with robotic sacrocolpopexy. MEASUREMENTS AND MAIN RESULTS: DNA was isolated and initially genotyped on a single nucleotide polymorphism (SNP) array to direct more detailed exome analyses. Exome sequences were mapped to the Human Genome Reference Sequence (GRCh37), and variants were compared between groups and to participants in the 1000 Genomes Project. Statistical analyses were performed using a software package commonly used in genetics research. TaqMan assay was used for verification, and p values were adjusted using the false discovery rate. Demographics of groups were compared using χ(2), Mann-Whitney U, and t tests. A SNP [rs171821] located near the ZFYVE16 gene was associated with patients but not control subjects, and the false discovery rate-adjusted p value was .046 (odds ratio, 45.2; 95% confidence interval, 5.06-403). Exome analyses of this gene yielded another SNP [rs249038 (G/A)] in 6 of 10 patients and none of the control subjects (p = .02). This SNP causes a heterozygous missense mutation of glycine to serine predicted to be deleterious by the Protein Variation Effect Analyzer and was also very rare among participants in the 1000 Genomes Project (p < .001). CONCLUSIONS: Two SNPs located near the ZFYVE16 gene on chromosome 5 may have played a role in the early, multicompartment sacrocolpopexy failure experienced by our patients. (www.clinicaltrials.gov Identifier: NCT01614587).

Alteration of apoptosis-related genes in postmenopausal women with uterine prolapse.

INTRODUCTION AND HYPOTHESIS: We aimed to compare expression levels of antiapoptotic and proapoptotic genes in parametrial and vaginal tissues from postmenopausal women with and without pelvic organ prolapse (POP). We hypothesized that the expression of genes that induce apoptosis may be altered in vaginal and parametrial tissues in postmenopausal women with POP. METHODS: Samples of vaginal and parametrial tissues were obtained from postmenopausal women with (n = 10) and without (n = 10) POP who underwent vaginal or abdominal hysterectomy. Expression levels of antiapoptotic (BCL-2, BCL-XL) and proapoptotic (BAX, BAD) genes were studied by real-time reverse-transcription polymerase chain reaction (RT-PCR). RESULTS: Gene expression levels of BCL-2 (P < 0.001), BCL-XL (P < 0.001), BAX (p = 0.001), and BAD (p = 0.004) were all higher in vaginal tissues from the POP group compared with the non-POP group. Similarly, gene expression levels of BCL-2 (p < 0.001), BCL-XL (p < 0.001), BAX (p < 0.001), and BAD (p < 0.001) in parametrial tissues were also significantly higher in the POP group compared with the non-POP group. Additionally, expression levels of BCL-2 (p = 0.05), BCL-XL (p < 0.05), BAX (p = 0.05), and BAD (p = 0.07) in the POP group were higher in parametrial tissue than in vaginal tissue samples. CONCLUSIONS: Antiapoptotic and proapoptotic gene expression levels differed significantly between postmenopausal women with and without POP. Bcl-2 family genes were overexpressed in the parametrium of patients with POP compared with vaginal tissue, suggesting that the processes responsible for POP have a greater effect on parametrial tissue than vaginal tissue during the development of POP.

Gene expression and immunoreactivity of elastolytic enzymes in the uterosacral ligaments from women with uterine prolapse.

Altered elastin metabolism has been documented in pelvic tissues from women with pelvic floor dysfunction. This study was conducted to quantify the expression of elastolytic enzymes in uterine cervix and uterosacral ligaments from women with uterine prolapse compared to asymptomatic normal controls. Paired tissues of uterosacral ligament and cervical tissues were obtained from 27 women with uterine prolapse and 14 normal controls. Steady state of matrix metalloproteinase 2 (MMP-2), tissue inhibitor of metalloproteinase 2 (TIMP-2), neutrophil elastase, α-1 antitrypsin immunoreactivity, and messenger RNA (mRNA) expression were analyzed by immunohistochemistry and real-time polymerase chain reaction, respectively. When compared with controls, women with uterine prolapse had a significantly greater level of MMP-2 immunoreactivity and mRNA expression, but less TIMP-2 and α-1 antitrypsin immunoreactivity and mRNA expression in their uterosacral ligaments. However, neutrophil elastase mRNA expression was similar between uterine prolapse and control tissue. Our results showed that there was a close relationship between expressions of MMP-2, TIMP-2, and α-1 antitrypsin in uterosacral ligament and the occurrence of uterine prolapse.

Histological, hormonal and biomolecular analysis of the pathogenesis of ovine Prolapsus vaginae ante partum.

The objectives of the present study were to evaluate the hormonal profiles, histology of the vagina and biomolecular analysis of connective tissue of ewes with and without vaginal prolapse. Blood samples from the jugular vein and biopsies of the vaginal tissue were taken from five late term pregnant, unaffected animals, four sheep during parturition and six ewes suffering from vaginal prolapse ante partum. The blood samples were submitted for determining the concentration of the steroid hormones progesterone by automatic luminescence immunoassay and estradiol-17ß by the sequence test. Investigations in the mRNA-expression including the estimation of the transcript levels of the α(2)-chain of collagen I, the collagenolytic metalloproteinase 1 (MMP 1), the tissue inhibitor of MMP 1 (TIMP 1) and the estrogen receptor α were carried out by using semiquantitative reverse transcription-PCR. Additionally, the histology of the vaginal wall of ewes with and without vaginal prolapse and animals intra partum was assessed. Because of a right-skewed distribution, data were logarithmised and described using the geometric mean (xg) and the dispersion factor (DF). The average progesterone concentration of affected ewes (xg = 19.35 ng/ml, DF 1.33) was above those of control animals ante (xg = 10.44 ng/ml, DF 1.58) and intra partum (xg = 9.24 ng/ml, DF 1.92). Compared to the pregnant control group (xg = 20.13 pg/ml, DF 1.49) the plasma levels of 17ß-estradiol in animals suffering from ante partum vaginal prolapse (xg = 27.81 pg/ml, DF 1.56) appeared to be slightly increased, but the difference was without statistical significance. The analysis of mRNA expression revealed a difference in the ante partum collagen metabolism in affected sheep. In prolapsed tissue the α2-chain of collagen I showed a decreased expression level in relation to the control animals in late-term pregnancy (P < 0.01). The average mRNA synthesis of MMP 1 or TIMP 1 in affected ewes was higher or lower, respectively, than the synthesis in healthy, late-term pregnant sheep. Significant differences were not observed. The production of transcripts of the estrogen receptor α was significantly decreased within the group of affected sheep compared to the unaffected pregnant ewes. Histological assessment showed that oedema was only detected in the subepithelial zone of the vaginal wall of intra partum sheep. There was no evidence for an inflammation of the prolapsed vaginal tissue since infiltration of leucocytes was present in all samples equally. The thickest vaginal epithelium due to hyperplasia of the epithelial cells was observed in sheep suffering from ante partum vaginal prolapse (xg = 83.95 µm, DF 1.21). This difference was statistically significant between the ante (xg = 31.12 µm, DF 1.22) and intra partum groups (xg = 33.27 µm, DF 1.24). Peripheral concentrations of progesterone and estradiol-17ß seem to have no influence on the occurrence of vaginal prolapse in ewes. Regarding histology of the vaginal wall in combination with the expression of local estrogen receptors, it was determined that there is neither a pronounced oedema nor an overexpression of the estrogen receptor α in affected animals, which means that no local estrogenic effect provokes the prolapse of vaginal tissue. The biomolecular analysis led to the new result, that ewes suffering from vaginal prolapse show alterations in the antepartal metabolism of vaginal connective tissue.

[MMP-1 and MMP-3 gene encoding polymorphism and the risk of the development of pelvic organ prolapse and stress urinary incontinence]. / Polimorfizm genów kodujacych MMP-1 oraz MMP-3 a ryzyko wystapienia wysilkowego nietrzymania moczu i zaburzen statyki dna miednicy mniejszej.

OBJECTIVES: The estimation of the association between the polymorphism at position -1607/1608 of the gene promoter encoding matrix metalloproteinase type 1 (MMP- 1) and the polymorphism at position -1612/1617 of the gene promoter encoding stromelysin type 1 (MMP-3) and the risk of the occurrence of pelvic organ prolapse (POP) and stress urinary incontinence (SUI). MATERIAL AND METHODS: 347 women were included into the analysis. POP study: the study group consisted of patients with clinically significant POP (POP-Q scale: 2, 3, 4). Women with normal pelvic floor statics (POP-Q scale: 0, 1) and not reporting symptoms of urinary incontinence were included into the control group. SUI study: the study group--patients with symptoms of stress urinary incontinence, the control group--continent women with normal pelvic floor statics (POP-Q scale: 0, 1). Samples of DNA were isolated from whole blood. The type of polymorphism was detected by RFLP method. RESULTS: Both, in the POP and the SUI study we have observed no statistically significant differences in the occurrences of MMP-1 and MMP-3 promoter polymorphisms between the study and the control groups. Also, the presence of the alleles G/GG (MMP-1) or 5A/6A (MMP-3) did not modify the risk of the POP and SUI development. CONCLUSIONS: Polymorphism type G/GG of gene promoter encoding MMP-1 and polymorphism type 5A/6A of the gene promoter encoding MMP-3 are not associated with the risk of the development of POP and SUI.

[Microarray analysis of gene expression profiles in pelvic organ prolapse].

OBJECTIVE: To identify the differentially expressed genes in cardinal ligament between patients with pelvic organ prolapse (POP) and postmenopausal women without POP by Human Genome Expression Chip and explore the potential molecular mechanism involved in POP. METHODS: From January to May, 2007, cardinal ligament samples were obtained from 3 postmenopausal patients with POP-Q stage III and 3 postmenopausal patients underwent hysterectomy due to other benign gynecologic diseases without POP in Peking Union Medical College Hospital. HE and Masson's trichrome staining was used to verify tissue origin and inspect histological changes. Those differentially expressed genes in cardinal ligaments were identified by Human Genome Chip and further interrogated with Gene Ontology (GO) and Pathway Analysis. Those remarkable expressed genes were confirmed by qRT-PCR. RESULTS: Alterations of ligament architecture in POP patients included disarrangement and collapse of smooth muscle bundles and collagen fibers. A total of 179 differentially expressed genes were screened between POP and non-POP cardinal ligament tissue, including 20 functional unknown genes. A total of 107 genes were upregulated in POP group, while 72 genes downregulated. Those differentially genes were revealed associated with multiple functional proteins and metabolic pathways by biological analysis. Among these, Wnt signaling pathway exhibited the most remarkable changes. Real-time quantitative PCR showed the genes of COL1A1, DKK1, SFRP1, FZD5, WNT16b in POP group (2.98+/-1.40, 3.03+/-0.48, 8.13+/-4.42, 5.19+/-3.50, 12.40+/-3.88) were upregulated significantly compared with non-POP group (1.09+/-0.08, 1.20+/-0.18, 0.41+/-0.51, 0.87+/-0.24, 1.40+/-0.47; P<0.05). CONCLUSIONS: The pathophysiology of POP is complex and associated with multiple functional proteins and metabolic pathways. Among these, the antagonist DKK1, SFRP1 in Wnt signaling pathway may contribute to a neurodegenerative role in POP development.

Differential expression of smooth muscle regulatory proteins in the uterosacral ligaments of women with uterine prolapse.

OBJECTIVE: To compare smooth muscle regulatory protein expression in the uterosacral ligament (USL) of women with and without uterine prolapse. STUDY DESIGN: USLs ligament were sampled in women with (n = 9) or without (n = 9) uterine prolapse. Caldesmon, smooth muscle actin (SMA), myosin heavy chain, and zinc finger protein messenger RNA expression was assessed by quantitative real-time polymerase chain reaction. Immunohistochemistry and digital image analysis were used to determine protein expression. RESULTS: Caldesmon messenger RNA expression and the ratio of caldesmon-SMA messenger RNA expression was significantly increased in the USL from women with uterine prolapse compared with women without prolapse (caldesmon mean +/- standard deviation messenger RNA, 0.81 +/- 0.46 vs 0.39 +/- 0.16; P = .01 and caldesmon-SMA messenger RNA ratio, mean +/- standard deviation, 0.11 +/- 0.04 vs 0.07 +/- 0.02; P = .01). In addition, the ratio of caldesmon-SMA staining was significantly increased in women with uterine prolapse compared with women without prolapse (mean +/- standard deviation, 0.44 +/- 0.28 vs 0.28 +/- 0.16; P = .03). CONCLUSION: Uterine prolapse is associated with an increased ratio of caldesmon-SMA actin expression.

Is laminin gamma-1 a candidate gene for advanced pelvic organ prolapse?

OBJECTIVE: We sought to determine allele frequencies of 3 LAMC1 single nucleotide polymorphisms (SNPs) in Caucasian and African American (AA) women with stage>II pelvic organ prolapse (POP) (cases) and in ethnicity-matched controls with stage

Differential expression of fibulins in the uterosacral ligaments of women with uterine prolapse.

PURPOSE: To compare fibulin-3 (FIB-3) and fibulin-5 (FIB-5) expressions in uterosacral ligaments (USL) of women with and without uterine prolapse. STUDY DESIGN: USL were sampled in a standardized fashion from women with (n = 8) or without (n = 8) uterine prolapse. Quantitative Real-Time PCR was performed to measure mRNA and immunohistochemistry to assess protein expression. RESULTS: FIB-3 mRNA expression and FIB-3 staining intensity was similar in the USL of women with and without uterine prolapse {[(FIB-3 mean +/- SD mRNA relative units) 0.45 +/- 0.41 vs. 0.46 +/- 0.82, NS] and [Intensity score, median (range), 1(0-1) vs. 1(0-1), NS]}. Both FIB-5 mRNA expression and FIB-5 staining intensity was significantly decreased in USL from women with uterine prolapse compared to women without prolapse {[(FIB-5 mean +/- SD mRNA relative units) 0.07 +/- 0.02 vs. 0.26 +/- 0.20, P = 0.02] and [Intensity score, median (range), 0(0-2) vs. 3(2-3), P = 0.002]}. CONCLUSION: FIB-5 expression is decreased in USL of women with uterine prolapse.

Extracellular matrix expression of human prolapsed vaginal wall.

OBJECTIVE: To compare the mRNA expression of extracellular matrix (ECM) proteins in postmenopausal prolapsed versus non-prolapsed anterior vaginal wall (AVW) tissue. We hypothesized that the weakening of the tissue leading to prolapse was due to decreased collagen production from a downregulation at the transcriptional level. METHODS: Following IRB approval, full thickness samples of redundant AVW were excised from consecutive age-equivalent, postmenopausal, women undergoing cystocele repair (prolapse, stage III or IV), or radical cystectomy (control, no clinical findings of prolapse). Total RNA was isolated, cDNA was synthesized, and quantitative real-time polymerase chain reaction (PCR) was conducted to assess the mRNA expression of collagens type I and III, pro-elastin, MMP3, MMP10, and MMP11. The significance of the difference of mRNA expression between prolapse and control tissues was tested using Student's t-test followed by Mann-Whitney Rank Sum Test. RESULTS: A 5.3-fold increase in collagen type I mRNA was found in prolapse (n = 47) over control (n = 7) tissues (P = 0.009). Type III collagen mRNA was also significantly increased to a 3.3 times higher level (P = 0.017). The ratio of type III to type I was decreased from 15.6 in controls to 9.7 in prolapse. An increasing trend in pro-elastin and MMP mRNA expression was found in prolapse, but this was not statistically significant. CONCLUSION: In this controlled study, the increase found in collagen mRNA expression disproved our hypothesis. To the contrary, this defective prolapsed tissue can signal its need for ECM replenishment. The message, however, is not being effectively translated to assist in tissue remodeling.

Does COLIA1 SP1-binding site polymorphism predispose women to pelvic organ prolapse?

INTRODUCTION AND HYPOTHESIS: COLIA1 polymorphism is associated with increased risk for stress urinary incontinence. We hypothesize that a similar association exists with pelvic organ prolapse (POP). METHODS: Patients with advanced prolapse and healthy controls were evaluated by interview, validated questionnaires, and pelvic examination. DNA was extracted from peripheral blood, and polymerase chain reaction was performed to determine the presence or absence of the polymorphism. Power calculation indicated the need for 36 patients in each arm. RESULTS: The prevalence of the polymorphic heterozygous genotype (GT) in the study and control groups was 33.3% and 19.4%, respectively, leading to an odds ratio of 1.75. This difference, however, did not reach statistical significance (p = 0.27). CONCLUSIONS: The COLIA1 polymorphism was not significantly associated with increased risk for POP.

COL3A1 2209G>A is a predictor of pelvic organ prolapse.

INTRODUCTION AND HYPOTHESIS: A familial tendency has been demonstrated in the etiology of pelvic organ prolapse (POP), but the specific genetic defects have not been identified. Type III collagen is an important factor in the repair of connective tissue, and gene polymorphisms may impair the tensile strength. We hypothesized that polymorphisms in the alpha I chain of the type III collagen protein-encoding gene (COL3A1) pose women at risk for POP. METHODS: In this case-control study, the prevalence of type III collagen polymorphisms was compared in women with and without signs and symptoms of POP. RESULTS: Two hundred and two POP patients and 102 normal parous controls were included. A homozygous single-nucleotide substitution in the coding region of type III collagen (COL3A1 2209G>A, rs1800255) was identified in 27 (13%) POP patients and three (3%) controls (odds ratio, 5.0; 95% confidence interval, 1.4-17.1). CONCLUSIONS: The probability of POP was higher in women with COL3A1 2209G>A. This polymorphism showed to be a relevant risk factor for POP.

Progesterone receptor polymorphism is associated with pelvic organ prolapse risk.

Progesterone and progesterone receptors (PGR) are known to play important roles in the pathophysiology of pelvic organ prolapse (POP). We investigated whether the PGR gene polymorphisms were associated with POP by conducting a case-control association study in 87 women with POP and 150 women without POP. Genotypes of the PGR gene polymorphisms (rs500760 and rs484389) were determined by polymerase chain reaction, followed by restriction fragment length polymorphism analysis. There was significant difference between women with and those without POP in the distribution of the PGR rs484389 genotypes evaluated. Using multivariable logistic regression, older age, increased body mass index, menopausal status, and PGR rs484389 genotype CT were significantly associated with POP. The present study shows that PGR genotype may be associated with POP.