Residual Disease In Patients Research Articles

Polymorphisms in the Multidrug Resistance Gene MDR1 (ABCB1) Predict for Molecular Resistance in Patients with Newly Diagnosed Chronic Myeloid Leukemia (CML) Receiving High-Dose Imatinib.

Abstract 2208Poster Board II-185 Introduction:Hematologic and cytogenetic responses to first-line imatinib are high in patients with chronic myeloid leukemia (CML), but only a minority of patients proceeds to a complete molecular response (CMR). Persistent residual disease at the molecular level may be designated as molecular resistance and has been suggested to be accounted for by quiescent malignant stem cells. P-glycoprotein (P-gp), which is encoded by the ABCB1 multi drug resistance (MDR1) gene, has been demonstrated to mediate efflux of imatinib. As hematopoietic progenitor cells efficiently express P-gp, it was hypothesized that single nucleotide polymorphisms (SNPs) of the ABCB1 gene that account for differences in functional activity may possibly be involved in the probability of developing a molecular response to imatinib and molecular resistance. Methods: We set out to evaluate whether the 3 most prevalent MDR1 SNPs (C1236T; G2677T/A; C3435T) would be associated with molecular response in a cohort of 46 early chronic phase CML patients who received 800 mg of imatinib. Molecular response was assessed according to the International Scale. Results: Patient characteristics, including age and Sokal risk score, were distributed evenly among groups of patients, classified according to SNP-genotype. The median follow-up was 46 months (range, 32-60 months). Each of the 3 SNPs were in Hardy-Weinberg equilibrium. However, each combination of 2 SNPs was in strong linkage disequilibrium (P<.001). The overall cumulative incidences of a major molecular response (MMR) and CMR were 78% and 41%, respectively, at 2 years. Molecular responses proved to depend strongly on SNP-genotype. A cumulative incidence of MMR of 52% and 50% was observed in patients with genotype 1236CT or TT as compared to 92% in patients homozygous for 1236C (hazard ratio's (HR): 0.32 and 0.33, P=.02) in univariate analysis, which remained significant (P=.03) following multivariate analysis. Patients homozygous for allele 3435T showed a CMR rate of 10% versus a CMR of 50% at 1 year in patients homozygous for allele 3435C (HR: 0.24 (0.07-0.83, P=.04). Hazard ratio's with respect to CMR associated with C1236T and G2677T were: 0.27 (0.08-0.97, P=.01) for the 1236TT genotype versus the 1236CC genotype, and 0.23 (0.06-0.88, P=.05) for the 2677TT genotype versus the 2677GG genotype, indicating a 4-fold reduction of the probability to develop a CMR. Given the strong linkage disequilibrium, the response differences could not unequivocally be attributed to a single SNP. However, given the enhanced P-gp activity earlier associated with the 2677TT genotype, it is suggested that enhanced clearance of imatinib by the 2677TT genotype may account for the lower incidences of a MMR and CMR in those patients. Conclusions: Molecular resistance in CML patients receiving high-dose imatinib appeared strongly associated with ABCB1 genes SNPs, suggesting a role for P-gp mediated drug efflux in malignant hematopoietic progenitor cells, that may possibly account for persistent molecular residual disease in patients favourably responding to imatinib. Disclosures:No relevant conflicts of interest to declare.

Flavopiridol in Chronic Lymphocytic Leukemia: A Concise Review

Patients with chronic lymphocytic leukemia (CLL) with high-risk cytogenetic features such as del(17p13) have limited treatment options and decreased overall survival. Dysfunction of p53 leads to resistance to fludarabine-based therapies. Cyclin-dependent kinase inhibitors (CDKi) are a novel class of agents that induce apoptosis in CLL cells independent of p53 mutational status. The synthetic flavone flavopiridol demonstrated promising in vitro activity in CLL. In initial phase I studies using a continuous infusion dosing schedule in a variety of malignancies, no clinical activity was observed. Detailed pharmacokinetic modeling led to the development of a novel dosing schedule designed to achieve target drug concentrations in vivo. In phase I testing, this dosing schedule resulted in acute tumor lysis syndrome (TLS) as the dose-limiting toxicity. With the implementation of a standardized protocol to prevent severe TLS, flavopiridol was administered safely, and responses were observed in heavily pretreated, fludarabine-refractory patients, cytogenetically high-risk patients, and patients with bulky lymphadenopathy. In a pharmacokinetic analysis, flavopiridol area under the plasma concentration-time curve (AUC) correlated with clinical response and cytokine release syndrome. Phase II studies are under way with encouraging preliminary results. Flavopiridol is currently under active investigation in combination with other agents and as a means to eradicate minimal residual disease in patients following cytoreductive chemotherapy. Several other investigational CDKi in preclinical and early clinical development are briefly discussed in this review.

Human Papillomavirus Test After Conization in Predicting Residual Disease in Subsequent Hysterectomy Specimens

To estimate the effectiveness of the human papillomavirus (HPV) test performed after conization in predicting residual disease in patients who subsequently underwent hysterectomy. A total of 115 patients who underwent hysterectomy after conization caused by cervical intraepithelial neoplasia grade 3 (CIN 3) and microinvasive cervical cancer (IA1 cancer) were included in this prospective study. All patients underwent HPV testing with a liquid hybridization assay immediately before hysterectomy. Differences in sensitivity, specificity, and accuracy between resection margin and the HPV test in predicting residual disease in subsequent hysterectomy samples were estimated using the McNemar exact test. Univariable analysis showed that age, parity, menopausal status, glandular extension, and severity of disease were not predictive for residual disease, but positive resection margin and positive HPV tests were significant factors for predicting residual disease. These factors were also significant in a multivariable analysis (positive resection margin 45.5%, odds ratio [OR] 3.09, 95% confidence interval [CI] 1.19-8.03, P=.021; positive HPV test 57.6%, OR 11.05, 95% CI 4.01-30.49, P<.001). With resection margin, the sensitivity, specificity, and accuracy in predicting residual disease were 75%, 53%, and 61%, respectively, whereas, with the HPV test, these values were 85%, 67%, and 73%, respectively (P=.454, .080, and .044, respectively). Of patients with positive resection margins, 79% of HPV-negative patients had no residual disease. Of patients with negative resection margins, no HPV-negative patient had residual disease. The HPV test after conization was significantly more accurate than resection margin for predicting residual disease. The predictive value of resection margin for predicting residual disease was much improved when used in combination with the HPV test. Use of the HPV test is recommended for identifying patients for subsequent hysterectomy after conization for CIN 3 and IA1 cancer. III.

A Rare e14a3 (b3a3) BCR-ABL Fusion Transcript in Chronic Myeloid Leukemia: Diagnostic Challenges in Clinical Laboratory Practice

Patients with chronic myelogenous leukemia have a t(9;22)(q34;q11.2) or variant translocation that results in a BCR-ABL fusion gene. BCR-ABL detection by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) is the standard practice for monitoring residual disease in patients with chronic myelogenous leukemia who receive tyrosine kinase inhibitor therapies. In this study, we describe a patient who tested positive for the BCR-ABL translocation by fluorescence in situ hybridization and cytogenetic analysis but tested negative by qRT-PCR molecular analysis at the time of diagnosis. Further PCR analysis and DNA sequencing with alternative primer sets demonstrated the presence of an e14a3 (also known as b3a3) BCR-ABL fusion. The e14a3 fusion is rare, but may be underreported as a result of many commercially available and laboratory-developed primer sets that fail to detect breakpoints in the ABL gene that are downstream of intron 1. For this patient, if the qRT-PCR assay had been used to monitor disease response/progression after treatment and not in conjunction with fluorescence in situ hybridization or cytogenetics at the time of diagnosis, the negative result would have been misinterpreted as molecular remission.

Significance of Increasing Levels of Minimal Residual Disease in Patients With Philadelphia Chromosome–Positive Chronic Myelogenous Leukemia in Complete Cytogenetic Response

The aim of this study was to evaluate the clinical relevance of increases in quantitative polymerase chain reaction (QPCR) levels in patients with chronic myelogenous leukemia (CML) who are in complete cytogenetic response (CGCR) on therapy. Patients with Philadelphia chromosome (Ph)-positive CML receiving tyrosine kinase inhibitors (TKIs) are frequently monitored for response by QPCR studies for minimal molecular disease. The clinical significance of increasing levels of QPCR in patients in CGCR is uncertain. One hundred sixteen patients in durable CGCR, and on imatinib therapy for at least 18 months, had increases in QPCR levels (documented at least twice consecutively) as defined by literature reports. These were further analyzed by the achievement of major molecular response (MMR) defined as QPCR < or = 0.05%, as well as by the degree of increase in QPCR. Only 11 (9.5%) of 116 patients with increases in QPCR had CML progression; 10 of them were among 44 patients (23%) who either lost a MMR or never had a MMR, and had more than 1 log increase of QPCR. Most patients with increases in QPCR remain in CGCR. Patients who lose a MMR or never achieve a MMR, and have more than 1 log increase of QPCR, should be monitored more closely, and may be evaluated for mutations of BCR-ABL kinase domain and considered for investigational therapeutic interventions.

The role of loop electrosurgical excisional procedure in the management of adenocarcinoma in situ of the uterine cervix

Objectives To evaluate the occurrence of residual or recurrent disease after loop electrosurgical excisional procedure (LEEP) for adenocarcinoma in situ (AIS) of the uterine cervix. Study design Records of 78 patients with a histological diagnosis of AIS of uterine cervix on LEEP who were treated and followed at our center between 1992 and 2008 were, retrospectively, reviewed. Results Of 78 patients who underwent LEEP, 47 had negative and 31 had positive resection margins. Of the 47 patients with negative margins, 30 underwent subsequent hysterectomy and residual AIS, including 1 invasive adenocarcinoma, was present in 17% (5/30) of patients. The remaining 17 had no additional procedures. Of the 31 patients with positive margins, 29 patients underwent subsequent hysterectomy and residual AIS, including 4 invasive adenocarcinomas, was present in 48% (14/29) of patients. The remaining two had no additional procedures. After a mean follow-up time of 28 months (range, 1–74 months), no recurrences were observed among the 19 patients who did not undergo hysterectomy. Conclusions The incidence of residual disease in patients with negative margins after LEEP for AIS of the uterine cervix is low but not negligible. Therefore, conservative management in these patients seems to be feasible but careful surveillance is required. However, positive resection margin carries a higher risk for residual AIS or occult invasive adenocarcinoma, warranting additional LEEP or hysterectomy in these patients.

Accuracy of DCE – MRI in predicting response in patients treated with taxane based neoadjuvant chemotherapy.

Abstract Abstract #5120 Background: Neoadjuvant chemotherapy for locally advanced breast cancer is an established treatment option. Several studies have shown DCE-MRI scanning to be the most accurate investigation to assess response to EC chemotherapy. Recent trials have shown that addition of Taxanes increases the overall response rate and the rate of breast conserving surgery. Taxanes act by inhibiting angiogenesis in the tumour and this could potentially affect the quality of the post treatment scan images as DCE-MR scan and cause inappropriate decisions regarding the required surgery.&amp;#x2028; Aim: To compare the accuracy of DCE-MR scan in identifying residual disease in patients treated with FEC (Fluoro uracil, Epirubicin, Cyclophosphamide) vs EC (Epirubicin, Cyclophosphamide) followed by Taxane.&amp;#x2028; Methods: Patients treated with primary chemotherapy who subsequently underwent breast conserving surgery were identified from a prospectively maintained database. Clinical details including chemotherapy regimes, pre and post chemotherapy MRI data, number of surgical procedures and histological parameters of the excised specimen were obtained.&amp;#x2028; Results: A total of 121 patients treated with primary chemotherapy were identified. Of these, 76 were treated with FEC or EC based chemotherapy and 45 were treated with EC+Taxane. DCE-MRI was significantly less accurate (p&amp;lt;0.05) when predicting the size of residual disease in the Taxane group.&amp;#x2028; &amp;#x2028; Conclusion: DCE MR scan is not as accurate for quantifying size of residual disease in patients treated with taxane when compared with patients treated with traditional FEC/EC based primary chemotherapy. This should be considered when planning surgery after neoadjuvant therapy. Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 5120.

Monitoring of minimal residual disease in patients with core binding factor acute myeloid leukemia and the impact of C-KIT, FLT3, and JAK2 mutations on clinical outcome

Mutational analysis of C-KIT, fms-like tyrosine kinase 3 (FLT3), and JAK2 genes was performed in 60 patients with core binding factor acute myeloid leukemia (CBF-AML). Patients reaching molecular remission had lower incidence of relapse and better overall survival (OS) than those not achieving molecular remission (p = 0.008 and 0.044, respectively). The overall incidence of C-KIT mutations was 33.3%, FLT3/internal tandem duplication (ITD) 6.6%, FLT3D835 10.0% and JAK2V617F mutations 3.3%. C-KIT mutations did not predict for clinical/molecular relapse (p = 0.33). OS of patients with C-KIT mutations was identical to patients without them when all patients with CBF-AML were analyzed together (p = 0.58). When AML1/ETO-positive patients were evaluated separately, OS in C-KIT-mutated patients was slightly inferior to unmutated ones (p = 0.14). Patients with CBF-AML with a mutated C-KIT gene were also more prone to extramedullary disease (p = 0.08). Of six patients harboring various FLT3D835 mutations, four (66.7%) relapsed, whereas among 43 cases without these mutations, 16 relapses (37%) were observed (p = 0.08). Our results on minimal residual disease, C-KIT, and FLT3/ITDs are in line with previous studies. Surprisingly, a possible role for FLT3D835 mutations was noted in addition. These results need validation in even larger patient cohorts than ours. For routine clinical practice, it may be meaningful to screen for C-KIT mutations in AML1/ETO-positive patients, as well as for FLT3D835 mutations in CBF-AML.

CTLA-4 genotype and relapse incidence in patients with acute myeloid leukemia in first complete remission after induction chemotherapy

The recently described single-nucleotide polymorphism CT60, located in the 3'-untranslated region of the CTLA4 (cytotoxic T-lymphocyte antigen 4 ) gene, has been associated with susceptibility to several autoimmune diseases and has also been shown to be involved in immune responses following allogeneic stem cell transplantation (SCT). However, the contribution of the CTLA4 genotype to the control of minimal residual disease in patients with acute myeloid leukemia (AML) has yet to be explored. We investigated the association between the CTLA4 CT60 A/G genotype and the incidence of leukemic relapse in 143 adult patients with AML in first complete remission after the same chemotherapy protocol (CETLAM LAM'03). The CT60 AA genotype was associated with a higher rate of leukemic relapse (56.4 vs 35.6%, P=0.004; hazard ratio (HR)=2.64, 95% confidence interval (CI)=1.36-5.14) and lower overall survival at 3 years (39.4 vs 68.4%, P=0.004; HR=2.80, 95% CI=1.39-5.64). This is the first study to report an association between polymorphisms at CTLA-4 and AML relapse.

Prognosticating of Relapse Risk Based on Multiparameter Flow Cytomeric Assessment of Minimal Residual Disease in Patients with Acute Myeloid Leukemia

Abstract Objective: To investigate the prognosticating of relapse risk based on assessment of minimal residual disease(MRD) in patients with acute myeloid leukemia(AML).Methods: Multiparameter flow cytometry (MPFC) analysis were used to detect the leukemia-associated aberrant immunophenotype (LAIP)of the pretreated patients with AML and assess the level of MRD after remission induction(Post-Ind MRD) and consolidation therapy(Post-Cons MRD).Results: The results showed that definite LAIP could be detected in about 94.3%of patients (115/122) with AML (except APL). Only one LAIP was identified in 15 cases (13.0%), but in other 100 cases (87.0%), two or more LAIP were identified. The most frequent LAIP identified was cross-lineage antigen expression (40.9%). The percentages of asynchronous antigen expression, antigen over-expression and antigen lack expression were 20.9%, 27.0% and 34.8% respectively. MRD frequency was monitored in 41 CR patients with AML after remission induction chemotherapy and 2 or more cycles of consolidation chemotherapy. 24 patients were Post-Ind MRD+ and 17 patients were Post-Ind MRD−. The percentages of relapse in Post- Ind MRD+ and Post-Ind MRD− were 75.0%(18/24) and 29.4%(5/17) respectively after consolidation chemotherapy. The relapsed free survival(RFS) of the patients with Post-Ind MRD+ and Post-Ind MRD− was 49.06±6.53 months and 11.92±1.64 months(P<0.0001) respectively. 18 patients were Post-Cons MRD+ and 23 patients were Post-Cons MRD−. The percentages of relapse in Post-Cons MRD+ and Post-Cons MRD− patients were 100%(18/18) and 21.7%(5/23) respectively after consolidation chemotherapy. The RFS of the patients with Post-Cons MRD+ and Post-Cons MRD− was 41.74±5.52 months and 10.06±1.72 months (P<0.0001) respectively.Conclusions: The level of post-Ind MRD and post-Cons MRD identified in the patients with AML was highly associated with their RFS. MRD measured by MPFC provided prognostic information in AML patients.

Dasatinib-Associated Major Molecular Responses Are Rapidly Achieved in Patients with Chronic Myeloid Leukemia in Chronic Phase (CML-CP) Following Resistance, Suboptimal Response, or Intolerance on Imatinib.

Dasatinib (SPRYCEL®) is a highly potent BCR-ABL inhibitor, with 325-fold higher potency than imatinib and a 16-fold higher potency than nilotinib in vitro. Across a series of phase II and III trials with more than 2 years of follow-up, dasatinib has demonstrated durable efficacy in patients with CML following resistance, suboptimal response, or intolerance to imatinib. Qualitative and quantitative assessment of BCR-ABL transcripts by RT-PCR is the most sensitive method for assessing minimal residual disease in patients with CML. In the treatment of patients with CML in the first-line setting, achievement of a major molecular response (MMR) within 18 months of therapy is considered to be clinically important. Here, molecular responses to dasatinib in patients with CML-CP following resistance, suboptimal response, or intolerance to imatinib were analyzed using data from two phase II studies (CA180-013 [START-C], -017 [START-R]) and a phase III dose-optimization study (CA180-034). BCR-ABL mRNA levels were determined in peripheral blood samples using real-time quantitative PCR and results were standardized using the international scale. A MMR was defined as a reduction of BCR-ABL transcripts to ≤ 0.1%. Samples were assayed at one of four laboratories (Mannheim, Germany; Adelaide, Australia; or Seattle, USA for −013 and −017; and Wallingford, USA for −034). Of 1,158 patients with CML-CP who were randomized to (017) or treated with (013/034) dasatinib, 1,067 had a molecular assessment and were included in the analysis. After 3, 6, 12, and 24 months of follow-up, a MMR was achieved by 12%, 22%, 35%, and 40%, respectively, of all patients analyzed. The 24-month MMR rate among patients who had achieved or maintained a complete cytogenetic response was 72%. In patients with imatinib resistance/suboptimal response (n=829), overall MMR rates after 3, 6, 12, and 24 months were 10%, 18%, 29%, and 34%, respectively, and in patients with imatinib intolerance (n=238), MMR rates were 22%, 37%, 55%, and 63%, respectively. In patients treated during the 034 study with dasatinib 100 mg once daily, which is associated with fewer key side effects, MMR rates at different time points were similar to rates with other dose schedules (24-month MMR: 100 mg QD, 36%; other schedules, 38%). Among all patients who achieved a MMR, median time to MMR was 5.7 months (6.0 months with 100 mg QD), and after 18 months, 92% of responding patients were without loss of MMR (88% with 100 mg QD). Overall, the results of this analysis demonstrate that high rates of MMR are rapidly achieved in patients with CML-CP treated with dasatinib following resistance, suboptimal response, or intolerance to imatinib.TableMajor molecular response (%)Follow-up (months)361224All analyzed patients (n=1067)12223540Resistant/suboptimal response (n=829)10182934Intolerant (n=238)22375563Phase II studies (013/017) (n=467)17273944Resistant (n=373)13213135Intolerant (n=94)33506978Phase III dose-optimization study (034) (n=600)9193238Resistant/suboptimal response (n=456)7162733Intolerant (n=144)15294654Response by dose schedule100 mg QD (n=154)718293670 mg BID (n=146)9183238140 mg QD (n=144)1322323850 mg BID (n=156)7173438

Significance of Rising Levels of Minimal Residual Disease in Patients with Philadelphia Chromosome-Positive Chronic Myelogenous Leukemia (Ph+ CML) in Complete Cytogenetic Response (CGCR)

Background. Patients with Ph+ CML receiving tyrosine kinase inhibitors (TKIs) are frequently monitored for response by quantitative polymerase chain reaction (QPCR) studies for minimal molecular disease. The clinical significance of rising levels of QPCR in CGCR is uncertain.Study Aims. To evaluate the relevance of increases of QPCR levels in patients with CML in CGCR on therapy.Study Group and Methods. Of 258 patients on imatinib therapy for newly diagnosed CML, 116 patients in durable CGCR on imatinib therapy for at least 18 months had significant QPCR increases (documented at least twice) as defined by literature reports. These were analyzed by the achievement of major molecular response (MMR; QPCR < 0.05%), and by the degree of QPCR increase.Results. The outcome of patients by disease status (still in MMR vs. loss of MMR vs. never in MMR) and by the QPCR level increase are shown in the Table. Only 13 of 116 patients (11%) with significant QPCR increases had CML progression; 11 of them were among 44 patients (25%) who either lost a MMR or never had a MMR, and had > 1 log increase of QPCR. The 5-year survival of all 116 patients was 92%, suggesting the minimal relevance of QPCR increases in patients in CGCR.Conclusion. Most patients with significant QPCR increases remain in CGCR. Patients who lose a MMR or never achieve a MMR, and have > 1 log increase of QPCR, should be monitored more closely, and may be evaluated for mutations of BCR-ABL kinase domain and considered for investigational therapeutic interventions. Allogeneic stem cell transplant should not be considered in view of the excellent survival.Outcome of Patients in CGCR by QPCR IncreasesDisease StatusQPCR Log increaseNo. PatientsCML ProgressionMedian follow-up from QPCR increase in months (range)Persistent MMRAny28036 (3–62)Loss of MMR>0.5–112034 (14–59)>1–225331 (6–52)>211445 (20–57)Not in MMR<132235 (10–70)>18425 (12–56)

Accuracy of magnetic resonance imaging in predicting residual disease in patients treated for stage IB2/II cervical carcinoma with chemoradiation therapy

The evaluation of residual disease (RD) after chemoradiation therapy (CRT) in stage IB2/II cervical carcinoma conventionally is based on a clinical examination and magnetic resonance imaging (MRI) performed 3 to 8 weeks after the end of treatment. Very few studies have correlated MRI and histologic findings specifically in cervix cancer. This was the objective of the current study. A retrospective review was undertaken of patients who fulfilled the following inclusion criteria: 1) stage IB2/II cervical cancer, 2) external radiotherapy (45 grays [Gy]) was given with concomitant chemotherapy followed by uterovaginal brachytherapy (15 Gy), 3) MRI studies were obtained between 3 weeks and 8 weeks after brachytherapy, and 4) completion surgery included at least a hysterectomy. Postsurgical histologic findings and MRI results were compared. Forty-four patients who were treated between 2003 and 2006 fulfilled all inclusion criteria. Twelve patients (27%) had achieved a radiologic complete response, 16 patients (36.5%) had unclassified lesions (RD or 'fibrosis'), and 16 patients (36.5%) had radiologic residual tumor. According to the histologic results, 19 patients (43%) had no RD, 10 patients (23%) had inframillimeter RD, 2 patients (5%) had RD that measured <1 cm, and 13 patients (29%) had RD that measured >1 cm. The sensitivity and specificity of MRI in evaluating RD were 80% and 55%, respectively. The current results indicated that the evaluation of RD 3 to 8 weeks after CRT with MRI is difficult, and the risk of false-positive results is high. Another radiologic procedure or a more technologically advanced MRI technique, such as diffusion-weighted MRI, should be evaluated.

A novel quantitative assessment of minimal residual disease in patients with acute myeloid leukemia carrying NPM1 (nucleophosmin) exon 12 mutations

Acute myeloid leukemia (AML) is a heterogeneous group of diseases affecting hematopoietic stem cells. In recent years, several novel molecular abnormalities have been identified in patients with AML, particularly in cases with normal karyotype. Among these, mutations of the nucleophosmin gene (NPM1) have been established as currently the most common abnormality in AML, found about half of all AML patients with normal karyotype. For the detection of NPM1 mutations on the molecular level, several different assays have been described, including direct sequencing, fragment analysis or high resolution melt analysis or high-performance liquid chromatography. NPM1 mutations might be also suitable as target structure for minimal residual disease monitoring. Here, we describe a novel quantitative assessment based on allelic discrimination assays and real-time PCR with mutation-specific minor groove binding (MGB) probes. This method offers an alternative to the standard routine laboratory evaluation and representing an efficient approach to the specific detection of NPM1 mutations without any false positivities caused by amplification on the wild-type alleles.

Adjuvant hysterectomy for treatment of residual disease in patients with cervical cancer treated with radiation therapy

The objective of this retrospective study was to determine the efficacy of adjuvant hysterectomy for treatment of residual disease in cervical carcinoma treated with radiation therapy. Between 1971 and 1996, 1590 patients with carcinoma of the uterine cervix (stages I–IIIb) were treated with radiation therapy. Three months after completion of radiation therapy, the status of local control was investigated, and total abdominal hysterectomy was performed in cases in which central residual disease existed in the cervix. Of the 1590 patients, residual disease was identified in 162 patients. Among these patients, 35 showed an absence of distant metastasis or lateral parametrial invasion and underwent hysterectomy. The overall 5- and 10-year survival rates for these patients were 68.6 and 65.7%, respectively. There was no significant difference in survival between patients with squamous cell carcinoma and those with non-squamous cell carcinoma or between patients with stage I/II carcinoma and those with stage III carcinoma. With respect to treatment-related morbidity, five (14.3%) patients suffered grade III or IV complications after hysterectomy. Adjuvant hysterectomy is an effective addition to radiation therapy in the treatment of cervical cancer, even in patients with stage III disease and in those with non-squamous cell carcinoma.

Rt-PCR method for diagnosis and follow-up of hematological malignancies: First approach in Bangladesh

Nested reverse-transcriptase polymerase chain reaction (rt-PCR) was performed on 58 leukemia patients at BIRDEM Laboratory, as a pioneering work in Bangladesh. Thirty of themwere examined for the presence of BCR-ABL being clinically and morphologically diagnosed as chronic myeloid leukemia (CML) and 28 for PML-RARalpha fusion transcripts being clinically and morphologically diagnosed as acute promyelocytic leukemia (APL/ AML M3). The cases were selected for targeted therapy with imatinib mesylate and all-Trans retinoic acid (ATRA) to treat CML and APL respectively. Samples were received either before commencement or during therapy. In the positive cases, amplified DNA products were visible after gel electrophoresis and were reported accordingly. In case of BCR-ABL, positive results were found for five out of six (83.33%) untreated cases and 11 out of 24 (45.83%) treated cases. Positive results for PML-RARalpha were found for 12 out of 14 (85.70%) untreated cases and 11 out of 16 (68.75%) treated cases. A strong positive correlation was found between duration of treatment and negativity of PCR results in both the cases. In present times, the detection of minimal residual disease in patients undergoing treatment for hematological malignancies has become an important goal, not only to monitor the effectiveness of therapy but also to detect an impending relapse. This is the first time in Bangladesh that rt-PCR method is being employed to detect or monitor the presence of abnormal fusion genes in hematological malignancies.

Blood Levels of Carbonic Anhydrase 9 Correlate with Clear Cell Renal Cell Carcinoma Activity

AbstractIntroductionBiomarkers for early detection of renal cell carcinoma (RCC) may help diagnose minimal residual disease in patients at risk for RCC, can guide anti-angiogenic therapy, or may help identify candidates for adjuvant treatment. In this study, we investigated whether blood levels of carbonic anhydrase 9 (CA9) correlate with RCC tumor burden and therefore disease activity.MethodsCA9 is a von Hippel–Lindau–hypoxia inducible factor target upregulated in clear cell RCC. We used an anti-CA9 antibody (M75)-based enzyme-linked immunosorbent assay test to measure CA9 levels in blood obtained before and after nephrectomy for clinically localized disease in patients with: (1) clear cell RCC, (2) papillary and chromophobe RCC or oncocytoma, or (3) benign kidney lesions, and we compared these samples to blood drawn from normal control individuals.ResultsWe observed a significant (p &lt; 0.006) decrease in the blood levels of CA9, after nephrectomy for localized disease, in the majority of patients with clear cell RCC (57%). In contrast, patients with nonclear cell RCC, benign disease, or those having undergone debulking nephrectomy for metastatic disease did not have a decrease in CA9 blood levels after nephrectomy. Preliminary longitudinal follow up measurements of CA9 levels in a small group of patients indicated that rising CA9 levels may correlate with disease progression.ConclusionsPlasma CA9 levels correlate with disease activity in a subset of clear cell RCC patients and should be considered in future multiplex RCC biomarker development algorithms.

Clinical significance of minimal residual disease in patients with t(8;21) acute myeloid leukemia in Japan

To examine the prognostic significance of minimal residual disease (MRD) in t(8;21) acute myeloid leukemia (AML), 96 bone marrow samples from 26 Japanese patients in complete remission (CR) were analyzed regarding the RUNX1/MTG8 transcript using real-time reverse transcriptase polymerase chain reaction assay. All patients were treated with intensive chemotherapy. The median copy number of the RUNX1/MTG8 transcript, measured after each treatment course decreased over time. However, an increase in the MRD level was documented in three patients after the second consolidation, and all of them subsequently relapsed. The relapse-free survival (RFS) did not differ between the patients whose MRD levels were below or above 1,000 copies/microg after the first consolidation, with respective 2-year rates of 62 and 86% (P = 0.21). With respect to the MRD level after induction therapy, our data also failed to show any favorable effect of a lower MRD on RFS. Although these findings need to be confirmed with a larger number of patients, our data indicate that the MRD level at a given time during the early course in CR does not predict the outcome in Japanese patients.

Accuracy of magnetic resonance imaging in predicting residual disease in patients treated for stage IB2/II cervical carcinoma with chemoradiation therapy: Correlation of radiological findings with histology of the cervix

16524 Background: The evaluation of residual disease (RD) after chemoradiation therapy (CRT) in stage IB2/II cervical carcinoma is conventionally based on clinical and radiological examinations (pa...

Monitoring minimal residual disease in patients with chronic myeloid leukemia after treatment with tyrosine kinase inhibitors

Monitoring for residual leukemic burden is an integral part of ongoing evaluation in chronic myeloid leukemia. With high rates of complete cytogenetic response induced by imatinib and other tyrosine kinase inhibitors, exploring refinements in current methods of minimal residual disease monitoring is relevant. Estimation of leukemic burden can influence treatment decisions. Real-time RT-PCR has become the method of choice for quantifying bcr-abl transcripts in patients undergoing therapy. Efforts are in progress to standardize this technique. The numbers of bcr-abl transcripts can vary in serial specimens, and it is important that changes in treatment (e.g. referral for transplantation, imatinib dose increase, or switch to a new tyrosine kinase inhibitor) be based on consistent rises in transcript number over time rather than on a single change. Prognosis in newly diagnosed patients with chronic myeloid leukemia in chronic phase has dramatically improved with the availability of imatinib and other tyrosine kinase inhibitors. In parallel, methods of monitoring minimal residual disease are evolving. Although real-time RT-PCR is now a standard monitoring method, the clinical significance of frequent and more precise follow up in patients achieving a complete cytogenetic response requires better definition.