Diagnosis of neuroblastoma and ganglioneuroma using Raman spectroscopy

Background

Raman spectroscopy has proven to be useful in studying premalignant and malignant lesions in adults. This is the first report to evaluate Raman spectroscopy in the diagnosis and classification of neuroblastoma in children.

Methods

A biopsy or resection of fresh tissue samples from normal adrenal glands, neuroblastomas, ganglioneuromas, nerve sheath tumors, and pheochromocytoma at our hospital were equally divided between routine histology and spectroscopic studies. At least 12 spectra were collected from different regions of each sample using a Renishaw Raman microscope. Raw spectra were processed to remove noise, florescence, and shot noise, and then analyzed using principle component analysis and discriminant function analysis.

Results

We collected 698 spectra from 16 neuroblastomas, 5 ganglioneuromas, 3 normal adrenal glands, 6 nerve sheath tumors, and 1 pheochromocytoma. Raman spectroscopy differentiated between normal adrenal gland, and neuroblastoma and ganglioneuroma with 100% sensitivity and 100% specificity. It correlated well with the Shimada histologic classification system with 100% sensitivity and 100% specificity. It was also able to differentiate neuroblastoma from nerve sheath tumors and pheochromocytoma with high sensitivity and specificity.

Conclusion

This technique can differentiate neuroblastoma from ganglioneuroma and other tumors. It has a potential as a noninvasive real-time diagnostic tool in classifying pediatric tumors.     

Differentiation of small round blue cell tumors using Raman spectroscopy

Purpose

Small round blue cell tumors (SRBCTs) are aggressive undifferentiated embryonal tumors, including neuroblastoma, rhabdomyosarcoma, Ewing sarcoma, and non-Hodgkin lymphoma. They share similar histologic features. Additional studies such as immunohistochemistry and molecular techniques are required to differentiate them. There is no widely available tool for real-time diagnosis. Raman spectroscopy is an analytical technique with potential for quick and accurate diagnosis of tumors in near real-time.

Methods

Fresh or banked frozen tissue samples from SRBCTs were processed for routine pathology and Raman spectroscopy. Raman results were correlated with the final pathology diagnosis.

Results

The data set was composed of 480 spectra from 32 samples, including 179 neuroblastoma, 37 Ewing sarcoma, 164 rhabdomyosarcoma, and 100 non-Hodgkin lymphoma spectra.Discriminant function analysis showed that a combination of 18 peaks could accurately identify 94% of spectra. At the tissue level, all tumors were correctly identified.Only 10 peaks were needed to classify all tissues with 100% accuracy. Spectral-level classification with this model was 87.9%.

Conclusion

Raman spectroscopy is an accurate technique for quickly and accurately differentiating SRBCTs. It could diagnose these specimens and provide a single, easy to use test for near real-time diagnosis.     

Characterization of pediatric Wilms' tumor using Raman and fluorescence spectroscopies

Purpose

Raman spectroscopy has been successfully demonstrated as an effective tool for tissue characterization and diagnosis, but nearly all studies have interrogated adult tissues and diseases. In this study, we demonstrate the application of Raman spectroscopy and its background autofluorescence for pediatric Wilms' tumor diagnosis.

Methods

Eight tumors were measured in this study, along with matched normal kidney tissue in 6 cases. Spectral comparisons were drawn, and diagnostic use was assessed using both the Raman spectral features as well as the inherent tissue fluorescence.

Results

The fluorescent background spectra were able to discriminate normal kidney from Wilms' tumor with 81% sensitivity and 100% specificity. The Raman spectra obtained 93% sensitivity and 100% specificity.

Conclusions

This pilot study shows that both autofluorescence and Raman spectra provide diagnostic use in discriminating Wilms' tumor from normal kidney. These techniques may be used individually or in tandem to develop a real-time intraoperative screening and diagnostic device.     

Wilms' tumor

In the last 2 decades, important advances in the treatment of Wilms' tumor have been made. The remarkable improvement in survival in these patients has been the product of new surgical techniques, classification of the tumors into prognostic stages upon initial presentation and the tailoring of chemotherapy and radiation therapy thus permitted. A brief historical perspective is presented with a review of the current treatment and ongoing studies.     

Wilms' tumor arising within a mediastinal teratoma

The majority of Wilms' tumors present in young children as an intraabdominal mass of renal origin. The use of multimodal treatment protocols according to internationally recognized treatment guidelines has resulted in extremely high cure rates even in the setting of advanced disease. By comparison, Wilms' tumors in older patients or Wilms' tumors located entirely outside the kidney are extremely rare. Because of the rarity of these tumors and the lack of established treatment guidelines specific to these lesions, older patients with atypical Wilms' tumors are at risk for under treatment and poorer outcomes. We report the case of an 18-year-old man with a Wilms' tumor arising within a mediastinal teratoma and discuss the management of this extremely rare lesion.     

Raman spectroscopy detects and distinguishes neuroblastoma and related tissues in fresh and (banked) frozen specimens

Background

Raman spectroscopy has been shown to accurately distinguish different neural crest-derived pediatric tumors. This study tests the ability of Raman spectroscopy to accurately identify cryopreserved tissue specimens using a classification algorithm designed from fresh tumor data and vice versa.

Methods

Fresh specimens of neuroblastoma and other pediatric neural crest tumors were analyzed with Raman spectroscopy. After analysis, the specimens were stored at −80°C. At a later date, the specimens were thawed and reanalyzed by Raman spectroscopy. A computer algorithm was used to classify the spectra from the frozen tissue against a computer model built on the fresh tissue data. This classification process was then reversed, testing fresh spectra against a model built from frozen data.

Results

We collected 1114 spectra (862 fresh and 252 frozen) from 62 tissue samples, including 8 normal adrenal glands, 29 neuroblastomas, 14 ganglioneuromas, 8 nerve sheath tumors, and 3 pheochromocytomas. At the tissue level, frozen neuroblastoma, ganglioneuroma, nerve sheath tumor, and pheochromocytoma were distinguished from normal adrenal tissue with 100% sensitivity and specificity. Fresh tissue had the same results except for the misclassification of one specimen of nerve sheath tumor.

Conclusions

The representative spectra show a high correlation between fresh and frozen tissue, and a clear difference between pathologic conditions. Spectra from frozen tissue can be accurately classified against spectra from fresh tissue and vice versa. This modality makes it possible to determine in a few minutes a result that often takes 12 to 36 hours for tissue processing and consideration by a trained pathologist to achieve.     

Bilateral intrarenal pelvis Wilms' tumor with fibroepithelial polyp

Intrarenal pelvis Wilms' tumor is rare in children. A case of a 28-month-old boy with bilateral intrarenal pelvis Wilms' tumor associated with a fibroepithelial polyp is reported in this article. The tumor was evaluated by ultrasonography, computed tomography, and intravenous pyelography. The boy underwent bilateral renal pelviotomies. Now he is being treated and followed up by pediatric oncologists.     

Teratoid Wilms' tumor: a case report with literature review

Teratoid Wilms' tumor is an unusual histologic variant of nephroblastoma in which the heterologous tissue predominates. The treatment for this tumor has not been established because of its rarity and varying tumor components. The authors report a case of localized teratoid Wilms' tumor in a 4-month-old female infant successfully treated with nephrectomy only. The tumor originated from the right kidney with multiple cysts and calcifications; renal teratoma was suspected preoperatively, and the operation was performed. The tumor was excised completely, and the specimen weighed 340 g. Histopathologic examination showed that various teratoid elements occupied most of the tumor with sparse islands of triphasic nephroblastoma, which was consistent with teratoid Wilms' tumor. Anaplastic elements, tumor capsule invasion, renal sinus soft tissue invasion, or tumors in intrarenal vessels were not observed. The patient was disease free for 3 years after surgery without receiving chemotherapy. We selected this therapeutic approach because of the chemoresistance of this tumor and the excellent prognosis in small (<550 g) stage I/favorable-histology classical Wilms' tumor diagnosed at less than 24 months of age. Therefore, nephrectomy alone may be an acceptable treatment of this stage of tumor in patients with teratoid Wilms' tumor.     

Teratoid Wilms' tumor: case report of a rare variant that can mimic aggressive biology during chemotherapy

Teratoid Wilms' tumor is considered by some as a variant of Wilms' tumor containing at least 50% heterologous differentiated tissue. Fewer than 30 cases have been described. We report a 9-month-old boy with bilateral Wilms' tumors who did not respond to multiagent chemotherapy and underwent right nephrectomy that showed a teratoid Wilms' tumor. The patient continued to survive despite cessation of treatment. The overall predominance of differentiated stromal elements in this subtype of Wilms' tumor might explain the poor response to chemotherapy yet generally favorable outcome. Recognition of this subtype on biopsy might justify earlier surgical intervention that, for bilateral tumors, might allow for greater nephron sparing.     

Ureteral extension in Wilms' tumor: a report from the National Wilms' Tumor Study Group (NWTSG)

Background

Extension of Wilms' tumor into the ureter is a rare event. We reviewed the National Wilms' Tumor Study Group (NWTSG) database to define the clinical presentation, associated pathologic features, and clinical outcome of these patients.

Methods

Records of children identified to have ureteral extension of Wilms' tumor enrolled in NWTS-3, 4, and 5 were reviewed. Presenting symptoms, diagnostic studies, histopathologic findings, operation performed, and outcome were recorded. The NWTS-5 surgical data were prospectively collected as part of the quality assurance program.

Results

Forty-five children were identified with ureteral extension. For NWTS-5, the incidence of ureteral extension was 2%. Clinical presentations were gross hematuria in 22 patients, 2 had passage of tissue per urethra, and 1 child had a urethral mass. The remainder had nonspecific presentations. Ureteral extension was seen on preoperative imaging in 14 patients, intraoperatively in 22, and on pathologic examination in 9 patients. Hydronephrosis was noted in 12 patients, and there was nonfunction of the kidney in another 8. Laterality of the tumor was right side in 26 and left in 19. Cystoscopy was performed in 12 children. Findings included tumor seen at the ureteral orifice in 6 patients and bleeding from the orifice in one child. All patients had radical nephrectomy including partial ureterectomy. The ureteral margin was positive in 3 patients, including 2 of the 7 with separate removal of the ureteral extension. The number of patients in each clinical stage was as follows: stage I, 10; stage II, 18; stage III, 14; and stage IV, 3. The tumor extended into the proximal ureter in 23 patients, distal ureter in 13, 7 had extension into the bladder, and 1 had urethral involvement. The level of ureteral extension was not clearly noted in the 45th child. The median follow-up was 96 months. Overall, 41 of 45 patients were alive at last contact. There were 3 deaths because of tumor in patients with unfavorable histologic tumors, and 1 because of treatment toxicity in a child with favorable histologic findings.

Conclusions

Ureteral extension occurs in approximately 2% of patients with Wilms' tumor. The diagnosis should be suspected in patients with gross hematuria, hydronephrosis, or nonfunctioning kidney. Cystoscopy with retrograde ureterogram may aid in preoperative diagnosis in these patients. Preoperative diagnosis is important because complete resection of the involved portion of ureter at the time of nephrectomy can avoid residual disease and the need for second surgery or radiation therapy.     

Analysis of near-infrared Raman spectroscopy as a new technique for a transcutaneous non-invasive diagnosis of blood components

. Near-infrared Raman spectroscopy can be a new technique for physical evaluations, allowing the measurement of lactic acid concentrations, in blood or muscles, during the physical activity in a transcutaneous non-invasive way. Lactic acid accumulation in the human body is one of the factors that leads to fatigue and therefore it should be continually monitored during physical training. Our proposal is to use Raman spectroscopy to monitor the lactic acid present in an athlete without interrupting his exercise for sample collection. The experimental set-up for Raman spectroscopy comprised a near infrared laser at 830 nm, a Kaiser f/1.8 spectrometer and a liquid nitrogen cooled CCD detector. The radiation from the exciting laser is blocked in the collecting system by Kaiser holographic filters. A personal computer controls the entire system, saving and processing the Raman spectra. Experiments were undertaken to verify the presence of lactic acid in the Raman spectra of solutions of lactic acid in human serum and in blood from a Wistar rat. After these two experiments, another was developed in vivo in a Wistar rat, injecting intraperitoneally 1 ml of a 0.12 mol/l lactic acid aqueous solution. An optical fibre catheter touching the skin of the rat groin, over the ileac vein collected the Raman signal. The presence of lactic acid was detected inside a live organism, in a transcutaneous non-invasive way. The minimum lactic acid concentration that the equipment can detect was also studied. An experiment was undertaken for that purpose, in which the laser illuminated directly a quartz cuvette containing solutions with decreasing lactic acid concentrations up to values near to the physiological level in the human body. The results indicated that the technique can be suitable for the physical evaluation of athletes. Paper received 12 May 2000; accepted after revision 29 June 2000.     

Isolated endobronchial metastasis of Wilms' tumor

Isolated endobronchial metastasis from extrapulmonary solid organ was rarely reported in previous literature. We report an isolated endobronchial metastasis of Wilms' tumor in a 4-year-old boy. He underwent right nephrectomy on account of Wilms' tumor at the age of 3 years. We performed a wedge resection of a metastasis to the right lower lobe of the lung a year later, followed by chemotherapy and irradiation of the involved lung over a period of 30 weeks. One and a half years later, he developed obstructive pneumonitis of the left lung. Fiberoptic bronchoscopy identified a tumor at the left main bronchus with near total bronchial obstruction, compatible with findings on a computed tomographic scan. Rigid bronchoscopy was performed to core out the obstructing tumor. It was reported as a metastatic Wilms' tumor. Successful reexpansion of the left upper lobe was achieved and he was discharged uneventfully, followed by adjuvant chemotherapy.     

Progress of fundamental research in Wilms' tumor

The progress of fundamental research on the histopathological and molecular genetic properties, model systems, growth factor involvement, and tumor markers of clinical nephroblastoma (Wilms' tumor) are reviewed. Histologically, Wilms' tumor (WT) has been found to reveal a disorganized renal developmental process in which blastema and epithelia are randomly interspersed in varying amounts of stroma. Anaplasia is the only criterion for assigning a WT as having an unfavorable histology. Cytogenetic analysis identified WT genes at chromosome 11p13 (WTI), 11p15 region (WT2), and 16q (WT3). Permanent in vitro WT cell lines and in vivo WT models, such as human xenografts, have been established which provide indefinite sources of tumor material for fundamental, as well as therapy-directed, research. Abnormalities of growth factor (GF) expression in WT indicate that GF may play an important role in WT pathogenesis. A series of monoclonal antibodies was tested in WT by immunohistochemical techniques to identify specific diagnostic and prognostic markers. p53 expression in anaplastic WT is significantly higher than in differentiated WTs, indicating p53 may be a prognostic marker. Although significant progress has been made in the fundamental research, our basic knowledge of this malignancy is still limited. The availability of suitable experimental models, particularly the human xenograft system, offers the opportunity for further study of the cell biological and molecular aspects of WT and its clinical progression.     

成人肾母细胞瘤的诊断与治疗(附9例报告及文献复习)

目的探讨成人肾母细胞瘤的诊断和治疗方法。方法回顾性分析9例成人肾母细胞瘤的临床表现、分期、治疗及预后。结果9例患者按照美国国家肾母细胞瘤研究组（National Wilms Tumor Study,NWTS）分期：Ⅰ期3例,Ⅱ期3例,Ⅲ期2例,Ⅳ期1例。9例均行肾切除术,其中术前分别辅以化疗1例、放疗1例、肾动脉介入栓塞化疗2例。9例术后辅以放疗和化疗。8例获得随访,随访1-6年,7例生存。结论成人肾母细胞瘤是一种比较少见的恶性肿瘤,早期诊断、手术治疗、辅以放疗和化疗等可明显提高治愈率并改善其预后。     

Laparoscopic nephroureterectomy for Wilms' tumor: oncologic considerations

Wilms' tumor is the most common malignant renal tumor of childhood. Surgical resection is an important aspect of therapy and is traditionally performed through an open transabdominal approach. The advent of advanced laparoscopic techniques for benign renal lesions has led to interest in applying a minimally invasive approach to Wilms' tumors in children. We describe a 2-year-old girl who presented with a right renal mass measuring 18 × 13 cm with peritoneal seeding and pulmonary metastatic disease. Wilms' tumor was confirmed on open biopsy. After neoadjuvant chemotherapy, a laparoscopic resection of the tumor with right radical nephroureterectomy, retroperitoneal lymph node dissection, and resection of peritoneal metastases was achieved using one 12-mm and four 5-mm ports. The specimens were removed in an endoscopic retrieval bag through a small Pfannenstiel incision by extending the 12-mm port site. The patient had an uncomplicated recovery and was discharged on the fifth post-operative day. She remains disease free at 19 months postoperatively. Minimally invasive techniques may be considered for resection of Wilms' tumor provided oncologic principles are carefully followed.     

Predictors of surgical outcome in Wilms' tumor: a single-institution comparative experience

Background

The merits of primary nephrectomy (PN) vs preoperative chemotherapy (PC) for patients with Wilms' tumor (WT) are much debated. Early data from the International Society of Pediatric Oncology suggested decreased intraoperative spillage but increased risk of local recurrence after PC. Patients with WT at our institution were managed with PC until 1996; subsequently, they underwent PN. This study compares these approaches as they affect tumor spillage, local recurrence, and survival.

Methods

Patients with WT diagnosed at the Hospital for Sick Children from 1985 to 2003 were reviewed.

Results

One hundred sixty patients were identified (114 PC and 46 PN). Tumor spill occurred in 6 (5.3%) of 114 PC and 2 (4.3%) of 46 PN patients. Tumor inhomogeneity, tumor size, and inferior vena cava compression/clot at diagnosis did not affect incidence of spill. Of 6 PC patients with surgical spill, 1 (17%) had significant tumor shrinkage, compared with 87 (81%) of 108 without spill (P < .001). Preoperative chemotherapy and PN had equal rates of surgical complications. Preoperative chemotherapy resulted in 12 (10.5%) of 114 local recurrences vs 5 (10.8%) of 46 with PN. Event-free survival and overall survival were 80% and 92% for PC at 129 months vs 85% and 96% for PN at 61 months.

Conclusions

Preoperative chemotherapy and PN are equally effective in the treatment of WT with no difference in tumor spillage. Failure of the tumor to shrink in size with PC was significantly associated with an increase in tumor spillage and would suggest that a more cautious surgical approach be undertaken in these cases.     

Adult Wilms' tumor: clinical characteristics of four cases and review of the literature

Adult Wilms' tumor (AWT) is a rare disease. We investigated the clinical characteristics and the outcome of four patients with AWT treated by nephrectomy followed by radiation therapy and/or chemotherapy (protocol of the Fourth National Wilms' Tumor Study). Serum levels of lactic dehydrogenase (LDH), immunosuppressive acidic protein (IAP), and C-reactive protein (CRP) were above normal in all four patients. Two of three patients with unfavorable histology (UFH) had stage III or IV disease and died within 14 months of diagnosis while undergoing the chemotherapy. The other patient with UFH had a stage II tumor, which was treated with tumor-bed irradiation, and has survived for 26 months after nephrectomy. One patient with a stage I tumor and favorable histology also received chemotherapy, but she subsequently developed multiple metastases, and died 70 months after nephrectomy. Radiotherapy for metastases had palliative effects on osseous pain and tumor growth. The clinical results of the chemotherapy for AWT were poor. Analysis of more cases is needed to indicate whether or not the conventional chemotherapeutic regimen for Wilms' tumor in children requires modification to improve the outcome in AWT. Received: February 22, 1999 / Accepted: July 23, 1999     

The use of preoperative chemotherapy in Wilms' tumor with contained retroperitoneal rupture

Purpose

The National Wilms Tumor Study currently describes 3 indications for the use of preoperative chemotherapy: extensive caval involvement, bilateral tumors, and patients who only have a single kidney. However, the management of patients who present with a contained retroperitoneal rupture is not specifically addressed. This is relevant because of the strong possibility of peritoneal contamination when performing a primary resection and the resultant requirement for total abdominal radiation. The use of neoadjuvant chemotherapy in this subgroup of patients may be warranted.

Methods

We retrospectively reviewed our experience with Wilms' tumor and identified 3 cases with contained rupture at presentation. Details of their initial evaluation and therapy, resection and pathologic findings, and follow-up constitute this report. Institutional review board waiver was obtained for the purposes of this review.

Results

Two male patients, aged 2.9 years, and 1 female patient, aged 9.3 years, were identified. All patients received preoperative chemotherapy with vincristine and dactinomycin (n = 1) plus doxorubicin (n = 2) for 4 to 6 weeks before surgical resection. One patient underwent pretreatment computed tomography-guided biopsy of the kidney mass for diagnostic purposes. Presurgical computed tomographic scans showed resolution of perinephric blood and fluid with tumor shrinkage. Histopathologic analyses showed all tumors were resected with negative margins, and there was no intraoperative tumor spillage. All patients received 1050 to 1080 cGy of flank radiation postoperatively. All patients are currently alive at follow-up without evidence of local recurrence or distant disease.

Conclusions

Neoadjuvant chemotherapy allowed for complete resection and avoidance of total abdominal radiation in 3 patients with ruptured Wilms' tumor and hematoma within the retroperitoneum. These data support the use of initial chemotherapy in children with retroperitoneal rupture and hematoma of Wilms' tumor at diagnosis.     

Case report of a Wilms' tumor and a left-sided inferior vena cava. A common tumor with a normal variant of surgical significance

We report a case of a successful surgical resection of a Wilms' tumor in the right kidney with a coincidental preoperative imaging finding of a left-sided inferior vena cava. To our knowledge, these 2 conditions occurring together has not been previously reported in literature. Diagnostic features and the value of magnetic resonance imaging are emphasized. We also review the literature of major venous anomalies and their influence on surgical procedures carried out on such patients.     

Raman spectroscopy for optical diagnosis in the larynx: preliminary findings

BACKGROUND AND OBJECTIVES: Raman spectroscopy (RS) provides information about molecular structure and is a potential tool for non-invasive tissue diagnosis. To determine if Raman spectra could be obtained rapidly from laryngeal tissue in vitro, and compare Raman spectra from normal, benign, and cancerous laryngeal tissue. STUDY DESIGN/MATERIALS AND METHODS: Forty-seven laryngeal specimens were studied using RS with signal acquisition times (SAT) between 1 and 30 second(s). Multivariate analysis was used to determine the diagnostic ability of RS compared to standard histology (n = 18, 13, and 16 respectively for normal tissue, carcinoma, and squamous papilloma). RESULTS: Good quality spectra were obtained with 5-second SAT. Spectral peak analysis showed prediction sensitivities of 89%, 69%, and 88%, and specificities of 86%, 94%, and 94% for normal tissue, carcinoma, and papilloma. CONCLUSIONS: In the larynx, spectral differences appear to exist between normal tissue, carcinoma, and papilloma. The ability to obtain spectra rapidly supports potential for future in vivo studies.