Ex vivo to in vivo model of malignant peripheral nerve sheath tumors for precision oncology.

BACKGROUND: Malignant peripheral nerve sheath tumors (MPNST) are aggressive soft tissue sarcomas that often develop in patients with neurofibromatosis type 1 (NF1). To address the critical need for novel therapeutics in MPNST, we aimed to establish an ex vivo 3D platform that accurately captured the genomic diversity of MPNST and could be utilized in a medium-throughput manner for drug screening studies to be validated in vivo using patient-derived xenografts (PDX). METHODS: Genomic analysis was performed on all PDX-tumor pairs. Selected PDX were harvested for assembly into 3D microtissues. Based on prior work in our labs, we evaluated drugs (trabectedin, olaparib, and mirdametinib) ex vivo and in vivo. For 3D microtissue studies, cell viability was the endpoint as assessed by Zeiss Axio Observer. For PDX drug studies, tumor volume was measured twice weekly. Bulk RNA sequencing was performed to identify pathways enriched in cells. RESULTS: We developed 13 NF1-associated MPNST-PDX and identified mutations or structural abnormalities in NF1 (100%), SUZ12 (85%), EED (15%), TP53 (15%), CDKN2A (85%), and chromosome 8 gain (77%). We successfully assembled PDX into 3D microtissues, categorized as robust (>90% viability at 48 h), good (>50%), or unusable (<50%). We evaluated drug response to "robust" or "good" microtissues, namely MN-2, JH-2-002, JH-2-079-c, and WU-225. Drug response ex vivo predicted drug response in vivo, and enhanced drug effects were observed in select models. CONCLUSIONS: These data support the successful establishment of a novel 3D platform for drug discovery and MPNST biology exploration in a system representative of the human condition.

Mutations involving TGFB and MAPK may be associated with malignancy in granular cell tumors.

Granular cell tumors (GrCTs) are mesenchymal neoplasms of presumed schwannian differentiation that may present as solitary or multifocal lesions with excision usually being curative. A minority of cases, however, show histological features associated with an increased risk for metastasis and are highly aggressive leading to death in about a third of cases. While benign and malignant cases have been shown to harbor mutations in the H + ATPase genes, there is only limited data examining molecular aberrations associated with malignancy. The departmental archives were searched for cases of atypical/malignant GrCTs. Clinical and histopathological features were noted. Whole-exome sequencing was performed. Three cases of malignant GrCTs and one case of atypical GrCTs were included. All three malignant tumors metastasized to distant sites with a median disease-free survival of 16 months and an overall follow-up time of 35 months. Whole-exome sequencing showed mutations involving TGFß and MAPK pathways in all four tumors. Although the cohort size is small, our preliminary findings suggest that mutations involving the TGFß and MAPK pathways may be associated with tumor progression or malignant transformation in GrCT pathogenesis.

Computational Genomics in the Era of Precision Medicine: Applications to Variant Analysis and Gene Therapy.

Rapid methodological advances in statistical and computational genomics have enabled researchers to better identify and interpret both rare and common variants responsible for complex human diseases. As we continue to see an expansion of these advances in the field, it is now imperative for researchers to understand the resources and methodologies available for various data types and study designs. In this review, we provide an overview of recent methods for identifying rare and common variants and understanding their roles in disease etiology. Additionally, we discuss the strategy, challenge, and promise of gene therapy. As computational and statistical approaches continue to improve, we will have an opportunity to translate human genetic findings into personalized health care.

Basal epithelial stem cells cross an alarmin checkpoint for postviral lung disease.

Epithelial cells are charged with protection at barrier sites, but whether this normally beneficial response might sometimes become dysfunctional still needs definition. Here, we recognized a pattern of imbalance marked by basal epithelial cell growth and differentiation that replaced normal airspaces in a mouse model of progressive postviral lung disease due to the Sendai virus. Single-cell and lineage-tracing technologies identified a distinct subset of basal epithelial stem cells (basal ESCs) that extended into gas-exchange tissue to form long-term bronchiolar-alveolar remodeling regions. Moreover, this cell subset was selectively expanded by crossing a cell-growth and survival checkpoint linked to the nuclear-localized alarmin IL-33 that was independent of IL-33 receptor signaling and instead connected to autocrine chromatin accessibility. This mechanism creates an activated stem-progenitor cell lineage with potential for physiological or pathological function. Thus, conditional loss of Il33 gene function in basal epithelial cells disrupted the homeostasis of the epithelial barrier at skin and gut sites but also markedly attenuated postviral disease in the lung based on the downregulation of remodeling and inflammation. Thus, we define a basal ESC strategy to deploy innate immune machinery that appears to overshoot the primordial goal of self-defense. Our findings reveal new targets to stratify and correct chronic and often deadly postviral disease.

Chromosome 8 gain is associated with high-grade transformation in MPNST.

One of the most common malignancies affecting adults with Neurofibromatosis type 1 (NF1) is the malignant peripheral nerve sheath tumor (MPNST), an aggressive and often fatal sarcoma that commonly arises from benign plexiform neurofibromas. Despite advances in our understanding of MPNST pathobiology, there are few effective therapeutic options, and no investigational agents have proven successful in clinical trials. To further understand the genomic heterogeneity of MPNST, and to generate a preclinical platform that encompasses this heterogeneity, we developed a collection of NF1-MPNST patient-derived xenografts (PDX). These PDX were compared with the primary tumors from which they were derived using copy number analysis, whole exome sequencing, and RNA sequencing. We identified chromosome 8 gain as a recurrent genomic event in MPNST and validated its occurrence by FISH in the PDX and parental tumors, in a validation cohort, and by single-cell sequencing in the PDX. Finally, we show that chromosome 8 gain is associated with inferior overall survival in soft-tissue sarcomas. These data suggest that chromosome 8 gain is a critical event in MPNST pathogenesis and may account for the aggressive nature and poor outcomes in this sarcoma subtype.

Comparison of the responsiveness of the SF-36, the Oxford Knee Score, and the Knee Society Clinical Rating System in patients undergoing total knee replacement.

OBJECTIVES: To compare the responsiveness of the Knee Society (KS) Clinical Rating System, the general health status measure Short Form 36 (SF-36), and both the raw and Rasch-based scores of the condition-specific Oxford Knee Score (OKS) in patients undergoing total knee replacement (TKR) METHODS: Data were prospectively collected as part of routine care from adult patients who underwent TKR between 2001 and 2006. OKS data fit the Rasch partial credit model after removing items regarding limping and kneeling. Responsiveness was assessed using effect size (ES), standardised response mean (SRM), and relative validity (RV). RESULTS: Among 702 patients with complete data at baseline and two follow-ups, the pain subscale of the KS (KS-P), raw-OKS, and Rasch-OKS consistently had higher levels of responsiveness than all eight SF-36 and the other KS subscales. At 6-month follow-up, Rasch-OKS had the largest ES and KS-P had the largest SRM (2.7 and 2.0, respectively). When compared to raw-OKS, the RVs of KS-P, Rasch-OKS, SF-36 bodily pain, and SF-36 physical functioning were 1.1, 0.66, 0.49, and 0.36, respectively. A similar ordering of responsiveness was observed at 24-month follow-up. CONCLUSION: The OKS and KS-P are more responsive than most SF-36 subscales in TKR patients. Raw-OKS and Rasch-OKS have comparable responsiveness. Different responsiveness indices may give different results.

Health-related quality of life after total knee replacement or unicompartmental knee arthroplasty in an urban asian population.

OBJECTIVE: To examine health-related quality of life (HRQoL) after total knee replacement (TKR) or unicompartmental knee arthroplasty (UKA). METHODS: Asian adult patients undergoing either TKR or UKA in a hospital in Singapore between 2001 and 2006 were interviewed before surgery and 6 and 24 months postoperatively to obtain demographic information and HRQoL scores using the Short Form-36 Health Survey (SF-36) and the Oxford Knee Score (OKS). RESULTS: Data were collected from 2243, 1715, and 1113 patients at baseline and at 6 and 24 months, respectively. TKR patients had a lower preoperative OKS than UKA patients and lower preoperative scores on four subscales of the SF-36 (P < 0.01). Both TKR and UKA patients' OKS and SF-36 subscale scores improved 6 months postoperatively except in the general health domain. SF-36 role physical and bodily pain scores showed the most improvement (40.9 and 33.0 points in TKR and 36.9 and 31.4 points in UKA patients, respectively). Two years after surgery, TKR patients' SF-36 scores and OKSs were not significantly different from those of UKA patients except for physical functioning scores. Multiple regression analysis adjusting for demographics showed that baseline scores were a significant predictor of the postoperative OKSs and scores on all SF-36 subscales (P < 0.01), whereas the type of surgery was not associated with the postoperative scores. CONCLUSIONS: Both TKR and UKA patients experienced significant improvements in HRQoL, particularly in the role physical and pain domains. After controlling for potential confounding variables, the type of surgery was not a significant predictor of patients' postoperative HRQoL scores.

A study on indirect and intangible costs for patients with knee osteoarthritis in Singapore.

OBJECTIVES: To estimate indirect costs through human capital approach and intangible costs through willingness-to-pay (WTP), and identify factors potentially affecting these costs in multiethnic Asian patients with knee osteoarthritis (OA). METHODS: Data were collected through face-to-face interviews among knee OA patients. Human capital approach was used to estimate indirect costs by multiplying: 1) days of absence from work because of OA, with average earnings per capita per day for working patients; or 2) productivity loss with the market price of housekeeping for retirees/homemakers. A closed-ended iterative bidding contingent valuation method was used to elicit willingness-to-pay for a hypothetical cure of OA as a proxy for intangible costs. Mann-Whitney U or Kruskal-Wallis H-tests were performed in univariate analyzes, and linear regression in multivariate analyses. RESULTS: Indirect costs per year and intangible costs were estimated at US$1008 and US$1200, accounting for 2.8% and 3.3% of annual household income, respectively. The indirect costs were significantly higher for male or working patients, while intangible costs were higher for Chinese, working patients, with higher income, or worse global well-being. CONCLUSION: This study demonstrated that eliciting indirect costs through human capital approach and intangible costs through WTP are acceptable and feasible in Asian patients with knee OA. Besides the direct costs, the indirect and intangible costs for the OA patients could be substantial.

Validation of Chinese Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) in patients scheduled for total knee replacement.

OBJECTIVE: Our aim was to cross-culturally validate Chinese Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) in patients with knee osteoarthritis (OA) scheduled for total knee replacement in Singapore. METHODS: Chinese WOMAC was translated from the original English version following standard guidelines. Patients were asked to complete a questionnaire containing the WOMAC (twice within 6 days), the Short Form 36 (SF-36), and the EuroQoL EQ-5D. Reliability was assessed using Cronbach's alpha and intraclass correlation coefficients (ICC), dimensionality using item-to-domain correlations and factor analyses. Convergent and discriminant construct validity was assessed using six each a priori hypotheses. RESULTS: Chinese WOMAC was well accepted by the patients in the pilot test. The Chinese and English versions were therefore administered to a consecutive sample of 131 Chinese- and 127 English-speaking subjects, respectively, with knee OA. Cronbach's alpha exceeded 0.7 for all domains except for Chinese pain domain, whereas the ICC exceeded 0.7 for all domains. Hypothesized item-to-domain correlations were observed for all items except for four items in Chinese physical function domain. The factor analyses yielded seven and five factors with eigenvalues of more than 1.0 in the Chinese and English versions, respectively. A total of 10/12 a priori construct validity hypotheses were satisfied for the Chinese version. Weak correlations between WOMAC pain and SF-36 bodily pain and moderate correlation between WOMAC pain and EQ-5D anxiety/depression were observed. CONCLUSIONS: Chinese WOMAC was well accepted and demonstrated acceptable psychometric properties in Singaporean patients with severe knee OA.

Computer-assisted minimally invasive total knee arthroplasty compared with standard total knee arthroplasty. A prospective, randomized study.

BACKGROUND: There is little information on the feasibility of computer navigation when using a minimally invasive approach for total knee arthroplasty, during which the anatomic landmarks for registration may be obscured. The purpose of the present study was to determine the radiographic accuracy of this technique and to compare the rate of functional recovery between patients who underwent computer-assisted minimally invasive arthroplasty and those who underwent conventional total knee arthroplasty. METHODS: One hundred and eight consecutive patients were randomized to undergo computer-assisted minimally invasive total knee arthroplasty or conventional total knee arthroplasty. Perioperative pain management was standardized. The clinical parameters, long-leg radiographs, and functional assessment scores were evaluated for six months postoperatively. RESULTS: Patients who underwent computer-assisted minimally invasive total knee arthroplasty had a significantly longer operative time (by a mean of twenty-four minutes) and a significantly shorter inpatient stay (3.3 compared with 4.5 days) in comparison with those who underwent conventional arthroplasty (p

Comparison of the EuroQol and short form 6D in Singapore multiethnic Asian knee osteoarthritis patients scheduled for total knee replacement.

OBJECTIVE: To compare the EuroQol (EQ-5D) and Short Form 6D (SF-6D) among multiethnic Asian patients with knee osteoarthritis (OA) scheduled for total knee replacement in Singapore. METHODS: Patients were asked to complete questionnaires including the EQ-5D, Short Form 36, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), and Lequesne knee index. EQ-5D and SF-6D utility scores were calculated using the scoring algorithms developed from the UK general population. Agreement between the 2 instruments was assessed by comparing their score distributions, means, medians, intraclass correlation coefficients (ICCs), and a Bland-Altman plot. Correlations of the EQ-5D and SF-6D with WOMAC and Lequesne knee index scores were also examined. RESULTS: A consecutive sample of 258 knee OA patients (127 English-speaking and 131 Chinese-speaking) participated. The mean +/- SD EQ-5D utility score was 0.49 +/- 0.31 (range -0.25-1.00) and the mean SF-6D utility score was 0.63 +/- 0.12 (range 0.32-0.89). In a hypothetical example, this 0.14-point difference in mean utility scores yielded a difference of $10,000/quality-adjusted life year (QALY) in cost-effectiveness ratios. The score distribution was bimodal for the EQ-5D and normal for the SF-6D. This poor agreement was also demonstrated by the Bland-Altman plot and the low ICC (range 0.18-0.54). Correlations of the WOMAC and Lequesne index with the EQ-5D were higher than with the SF-6D. CONCLUSION: Using different preference-based health-related quality of life instruments may yield different utility scores, which could have a great impact on QALY estimates. This highlights the importance of selecting appropriate instruments for economic evaluation. Additional research is needed to determine which instrument (the EQ-5D or the SF-6D) should be used in OA patients.

Direct and indirect costs of osteoarthritis in Singapore: a comparative study among multiethnic Asian patients with osteoarthritis.

OBJECTIVE: To estimate and compare the direct and indirect costs of osteoarthritis (OA) in multiethnic Asian patients with OA in Singapore. METHODS: The study was a retrospective and cross-sectional design. Patients were stratified according to ethnicity and presence or absence of joint surgery. Direct costs were estimated from both a societal and a patient perspective using the Singapore General Hospital database; indirect costs were estimated using the human capital approach. All costs were expressed as mean costs per patient per annum in 2003 Singapore dollars. RESULTS: A total of 1179 patients (83.6% Chinese, 7.2% Malay, 3.5% Indian, 5.7% others) were included in estimating direct costs, of which 513 (43.5%) had total knee replacement (TKR) and 92 (7.8%) total hip replacement (THR), while 105 patients (71.4% Chinese, 14.3% Malay, 14.3% Indian) were included in estimating indirect costs. Direct costs to patients ranged from 1460 dollars to 7477 dollars for Chinese, 1362 dollars-7211 dollars for Malays, 1688 dollars-6226 dollars for Indians, and 1437 dollars-12,140 dollars for other ethnic patients; direct costs to society ranged from 3351 dollars to 15,799 dollars for Chinese, 2939 dollars-15,436 dollars for Malays, 3150 dollars-10,990 dollars for Indians, and 2597 dollars-17,879 dollars for other ethnic patients. In contrast, the indirect costs ranged from 1215 dollars to 3834 dollars for Chinese, 1138 dollars-6116 dollars for Malays, and 1371 dollars-5292 dollars for Indians. However, most ethnic variations were not statistically significant. CONCLUSION: The economic burden of OA to society and patients increased by 3-fold or more in the patients with TKR/THR compared to those without. The ethnic differences in health resources consumed were more apparent when the disease progressed.

Stress fracture of the medial tibial plateau after minimally invasive unicompartmental knee arthroplasty: a report of 2 cases.

Minimally invasive unicompartmental knee arthroplasty has been increasingly popular for the past few years. Several short- and medium-term published reports have shown it to be as efficacious as open unicompartmental knee arthroplasty, with reduced morbidity. However, complications specifically related to the minimally invasive techniques are not well documented. We report 2 cases of stress fracture of the medial tibial plateau after unicompartmental knee arthroplasty with minimally invasive instrumentation. Awareness of this as a possible complication, with increased care in surgical technique, will help to avoid this problem.

Isolated congenital pseudoarthrosis of the fibula.

Congenital pseudarthrosis of the limb most commonly involves the tibia, although various combinations of bones including fibula, radius, ulna, clavicle and humerus have all been described. Isolated congenital pseudarthrosis of the fibula is a very rare entity with only 12 cases reported in the English literature. We report three cases of this condition treated in our institution. The first child had a varus ankle deformity at the age of 4 months. The other two children presented with valgus ankle deformity after they started to walk. Two patients were treated conservatively while the third had a distal tibio-fibular fusion in view of severe valgus deformity. All three patients showed good early results after 1 to 2 years. We advocate early distal tibio-fibular fusion to prevent valgus deformity in these children.