In multiple myeloma, t(4;14)(p16;q32) is an adverse prognostic factor irrespective of FGFR3 expression

Blood ◽  
2003 ◽  
Vol 101 (4) ◽  
pp. 1520-1529 ◽  
Author(s):  
Jonathan J. Keats ◽  
Tony Reiman ◽  
Christopher A. Maxwell ◽  
Brian J. Taylor ◽  
Loree M. Larratt ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Growth Factor Receptor ◽  
Poor Response ◽  
Response To Therapy ◽  
Prognostic Indicators ◽  
Rt Pcr ◽  
Poor Survival ◽  
Amino Terminal ◽  
Breakpoint Analysis ◽  
Polymerase Chain

This study analyzed the frequency and clinical significance of t(4;14)(p16;q32) in multiple myeloma (MM) among 208 patients with MM and 52 patients with monoclonal gammopathy of undetermined significance (MGUS); diagnosed between 1994 and 2001. Patients with the translocation were identified using reverse transcription–polymerase chain reaction (RT-PCR) to detect hybrid immunoglobulin heavy chain (IgH)–MMSET transcripts from the der(4) chromosome. We found 31 (14.9%) t(4;14)+ MM patients and 1 (1.9%) t(4;14)+ MGUS patient. IgH-MMSET hybrid transcripts were detected in bone marrow (BM) and blood. Breakpoint analysis revealed that 67.7% of t(4;14)+ patients expressed hybrid transcripts potentially encoding full-length MMSET, whereas the remainder lacked one or more amino terminal exons. Expression of fibroblast growth factor receptor 3 (FGFR3), presumptively dysregulated on der(14), was detected by RT-PCR in only 23 of 31 (74%) patients with t(4;14)+ MM. Patients lacking FGFR3 expression also lacked detectable der(14) products. Longitudinal analysis of 53 MM patients with multiple BM and blood samples showed that, over time, BM from t(4;14)+ patients remained positive and that t(4;14)− patients did not acquire the translocation. IgH-MMSET hybrid transcripts and FGFR3 transcripts disappeared from blood during response to therapy. No correlation was observed between the occurrence of t(4;14) and known prognostic indicators. However, we find the t(4;14) translocation predicts for poor survival (P = .006; median, 644 days vs 1288 days; hazard ratio [HR], 2.0), even in FGFR3 nonexpressors (P = .003). The presence of t(4;14) is also predictive of poor response to first-line chemotherapy (P = .05). These results indicate a significant clinical impact of the t(4;14) translocation in MM that is independent of FGFR3 expression.

scholarly journals A reverse transcriptase-polymerase chain reaction detects heterogeneous chimeric mRNAs in leukemias with 11q23 abnormalities

Blood ◽  
1994 ◽  
Vol 83 (10) ◽  
pp. 2912-2921 ◽  
Author(s):  
K Yamamoto ◽  
M Seto ◽  
S Iida ◽  
H Komatsu ◽  
N Kamada ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Response To Therapy ◽  
Chromosome 9 ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Single Clone ◽  
Sensitivity Studies ◽  
Polymerase Chain ◽  
11Q23 Abnormalities

Abstract The MLL gene involved in 11q23 translocations found in the majority of infantile leukemias and some secondary leukemias makes fusion transcripts with genes such as LTG4 (chromosome 4), LTG9 (chromosome 9), and LTG19 (chromosome 19) as a result of reciprocal translocation. We have examined 25 cases of leukemias with 11q23 abnormalities by Southern blot analysis and the reverse transcriptase-polymerase chain reaction (RT-PCR). Using various primer pairs, chimeric mRNAs could be amplified in 6 of 7 leukemias with t(4;11), 6 of 8 leukemias with t(9;11) including secondary leukemia, 8 of 9 leukemias with t(11;19), and 1 with a deletion at 11q23. The chimeric mRNAs were heterogeneous and differential usage of the MLL exons was found, irrespective of the partner chromosomes. Sensitivity studies showed that a single clone with chimeric mRNA in 10(4) to 10(5) cells could be detected. These findings show that the present RT-PCR settings provide a rapid, accurate, and sensitive tool for diagnosing leukemias with 11q23 translocations and for monitoring response to therapy in these patients.

The Platelet-Derived Growth Factor Receptor beta Fuses to Two Distinct Loci at 3p21 in Imatinib Responsive Chronic Eosinophilic Leukemia.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3253-3253 ◽  
Author(s):  
Claire Curtis ◽  
Jane F. Apperley ◽  
Raymond Dang ◽  
Michael Jeng ◽  
Jason Gotlib ◽  
...  
Keyword(s):  
Amino Acid ◽  
Tyrosine Kinase ◽  
Blast Crisis ◽  
Coiled Coil ◽  
Growth Factor Receptor ◽  
Myeloproliferative Disorders ◽  
Rt Pcr ◽  
Chronic Eosinophilic Leukemia ◽  
Amino Terminal ◽  
Exon 11

Abstract We have identified three patients (2 adults, one infant) who presented with BCR-ABL negative eosinophilic myeloproliferative disorders. Cytogenetic analysis revealed a t(1;3;5)(p36;p21;q33) for case 1 and a t(3;5)(p21–25;q31–35) for cases 2 and 3. Two-color fluorescence in situ hybridization (FISH) using differentially labelled probes flanking PDGFRB indicated that this gene was disrupted in all three cases. 5′ rapid amplification of cDNA ends (5′RACE) for case 1 identified an in-frame mRNA fusion of exon 9 of the WDR48 gene at 3p21 to exon 12 of PDGFRB. The chimeric mRNA is predicted to encode a 872 amino acid fusion protein that retains the amino terminal WD repeat region of WDR48 fused to the transmembrane and intracellular tyrosine kinase domains of PDGFRbeta. Cases 2 and 3 were negative for the WDR48-PDGFRB fusion mRNA by RT-PCR using several combinations of primers. 5′RACE PCR from case 2 RNA identified a fusion involving a second 3p21 gene: GOLGA4 exon 11 was fused in-frame to exon 11 of PDGFRB. Exactly the same fusion was found in case 3. The predicted 991 amino acid protein included the amino terminal coiled-coil domain of GOLGA4 fused to the transmembrane and intracellular tyrosine kinase domains of PDGFRbeta. Interestingly, both WDR48 and GOLGA4 are involved in endocytic shuttling pathways. The presence of all fusions was confirmed by RT-PCR and identification of the genomic breakpoints. Imatinib, a known inhibitor of PDGFRbeta, selectively blocked the growth of patient CFU-GM for case 2. Following the identification of PDGFRB rearrangements, all three patients were treated with imatinib. Case 1 was in transformation, but responded rapidly to minimal doses of imatinib (800mg daily for 4 days) with complete cytogenetic remission but remained pancytopenic. Blast crisis recurred 8 months later, responded similarly to 3 days of imatinib, but the patient died 2 months later of invasive fungal infection. Case 2 responded clinically and remains in sustained cytogenetic and molecular remission (nested RT-PCR negative for GOLGA4-PDGFRB). Case 3 (a 13 month old boy) had a complete hematologic response to 50mg/day imatinib but the t(3;5) was still seen in 40% of metaphases at 3 months. We conclude that PDGFRB fuses to diverse partner genes to give rise to atypical MPDs. Although very rare, identification of these fusions is critical for proper management of affected individuals.

IgM Multiple Myeloma: A Rare Subtype Highly Sensitive to Lenalidomide

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 5220-5220
Author(s):  
Alvaro Moreno-Aspitia ◽  
Antony Charles ◽  
Tejal Patel ◽  
Celine Bueno ◽  
Abba Zubair ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Transient Response ◽  
Low Dose ◽  
Plasma Cells ◽  
Poor Response ◽  
Response To Therapy ◽  
Bone Lesions ◽  
Best Response ◽  
M Spike

Abstract Background: IgM multiple myeloma (MM) are very rare plasmaproliferative disorders representing 0.5–1.2% of all cases of MM and < 0.2% of all IgM monoclonal gammopathies. Clinical criterion are not always helpful in differentiating IgM MM from Waldenstrom macroglobulinemia. However, the presence of lytic bone lesions, absence of lymphadenopathy and/or hepatosplenomegaly, presence of translocation of the immunoglobulin heavy chain locus at 14q32 [t(11;14), t(14;16), t(4;14)], and strong expression of CD138 by the plasma cells are useful in the diagnosis of IgM MM. It has been our experience and of others that these cases have an aggressive behavior at presentation, shorter survival than IgG and IgA MM and poor response to therapy for lymphoplasmacytoid lymphomas. We present here 2 cases of IgM MM with a dramatic response to Lenalidomide and low dose dexamethasone (Rev/Dex) Results: Baseline patient characteristics at time of diagnosis of IgM MM and therapy outcome are presented in the following 2 tables: Table 1. Case 1 2 Age and sex 72 (F) 73 (F) Serum M-spike (g/dL) 5.3 6.2 Urine M-spike (mg/dl/24 hrs) 72 412 Serum IgM (mg/dL) 8,590 11,000 BM plasma cells percentage 90 20 Plasma cell immunophenotyping CD138+++, partial CD20, CD56− CD138+++, partial CD20, CD56− Cytogenetics (Standard and/or FISH) Standard: normal FISH: not done on initial biopsy. On follow up there were insufficient number of plasma cells to perform test Standard: of 20 metaphases, 6 had a complex hypotetraploid karyotype with relative loss of 13q, 14, 15, 16, 20, and 22, and numerous unbalanced rearrangements. FISH: a plasma cell clone with monosomy 13 and IGH/c-MAF fusion, t(14;16). In addition, approximately 60% of plasma cells had a tetraploid clone with the same anomalies as well as relative loss of p53 Bone lesions Multiple non-traumatic spinal fractures and of stenum Several lytic lesions of long bones Renal insufficiency No No Anemia (Hbg g/dL) Yes (8.7) Yes (8.1) Hypercalcemia (Ca mg/dL) Yes (12.5) Yes (11.4) Beta 2 microglobulin (mg/dL) 5.79 8.51 Serum viscosity (cpoise) 5.9 4.8 Table 2. Best Response to therapy Case Therapy Best Response Comments 1 Rituxan, then Fludarabine based therapy Transient response Rapid progression after partial and transient response to each therapy 1 Lenalidomide + LD-Dex sCR after cycle #6. Currently on CR 18 months later IgM declined from 8,590 to 43 mg/dL after 4 cycles of Rev/Dex. 2 Lenalidomide + LD-Dex VGPR after cycle #2 IgM declined from 11,000 to 463 mg/dL after cycle 3. Complete disappearance of M-spike in serum; BM to be done after cycle #4 Conclusions: This is the first report that we are aware of a rapid and dramatic response to lenalidomide and low dose dexamethasone in these rare cases of IgM MM with poor response to NHL-type treatment. Lenalidomide-based therapy might abrogate poor prognosis cytogenetics in this unusual subtype of MM (case #2), however, follow up for this patient is still very short.

Identification of Genes Associated with Resistance and Response in Vivo to Therapy with Rituximab, Fludarabine and Cyclophosphamide in Patients with Chronic Lymphocytic Leukemia.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1622-1622 ◽  
Author(s):  
Medhat Shehata ◽  
Dita Demirtas ◽  
Stefanie Tauber ◽  
Susanne Schnabl ◽  
Martin Bilban ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Expression Signature ◽  
Poor Response ◽  
Response To Therapy ◽  
Lymphocytic Leukemia ◽  
Pcr Analysis ◽  
Poor Responders ◽  
Rt Pcr ◽  
Number Of Patients

Abstract Fludarabine and cyclophosphamide are the backbone of therapy for patients with CLL. The addition of rituximab leads to further improvement of response rates and progression free survival (results from the randomized CLL8 study of the GCLLSG). However, a considerable number of patients have insufficient or short responses to FC or RFC and there is a need to identify factors influencing response or resistance to therapy. The aims of this study are to identify gene expression associated with response or resistance before the start of therapy, to investigate changes in the expression of specific genes or pathways associated with response or resistance during the first cycle of FC and RFC, to provide a rationale for the additional use of novel drugs to improve remission and overcome resistance. We investigated peripheral blood samples from 20 patients receiving FC (n=10) or RFC (n=10) by gene expression profiling, flow cytometry, RT-PCR and western blotting before and during therapy. Sixteen patients received FC or RFC as first line (8 within the CLL8 study) and 4 as second line treatment. All patients were in stage Binet B or C. Gene expression was analyzed and correlated to good (CR or PR) or poor clinical response (SD or PD) at the end of therapy based on NCI-WG/IWCLL criteria. CD19+ cells were harvested by cell sorting before therapy, 24 hours after FC (FC arm), 24 hours after rituximab, and 24 hours after FC (RFC arm). Microarray analysis was performed using Affymetrix U133A gene chips. Genes with a consistent pattern of expression (high or low) in the majority of samples in the good or poor response group were further analyzed. Overall, 9 patients responded adequately to therapy (3 CR, 7 PR), while 11 did not (7 SD, 3PD). Unmutated IgVH status and poor risk cytogenetics were more frequent in poor responders. Gene expression signature before treatment showed that overexpression of 39 genes strongly correlated with response, while overexpression of 20 genes (including HSPA1B, IFI6, APP, CEACAM1, CD9, GAB1, INPP5F) was associated with resistance. Changes in expression after initiation of treatment was also analyzed. Seven genes (including CENTD1, HBA2, COL9A2 and APRIN) were significantly upregulated after rituximab in non-responders. Upregulation of 13 genes (including PMAIP1, SFRS11, CLK1, EFHC1, MRPL39, TUG1, TBRG1, CD49d, PTPRC) after R-FC and 7 genes (including ITPKB, LOC641298, CD44, TAF5) after FC was associated with poor response (resistance) to RFC and FC respectively. Many of these genes are involved in regulation of apoptosis, cell cycle, integrin and PI3-K signaling Therapeutic antibodies or inhibitors against some of these targets are already available. RT-PCR analysis demonstrated a significant downregulation of Akt1 mRNA 24 hours after rituximab infusion in RFC group but no significant changes were observed in patients receiving FC alone. In vitro exposure to rituximab confirmed its in vivo effect and resulted in a significant downregulation of Akt1 and PI3-K-p85 mRNA expression. FACS analysis demonstrated a decrease in the percentage and mean fluorescence intensity (MFI) of surface CD20 after rituximab infusion. This effect was associated with a significant change in total amount and phosphorylation state of CD20 in the RFC group. There was also a decrease in the MFI of CD44 and CD23 after rituximab in the majority of patients in the RFC group but this effect was not consistent in the FC group. In conclusion, we have identified a set of markers associated with good or poor response to FC or RFC before therapy and during the first cycle of treatment. The data provide a rationale for targeted drug combinations to overcome resistance and improve response to therapy in CLL.

Human Epidermal Growth Factor Receptor 2 Assessment in a Case-Control Study: Comparison of Fluorescence In Situ Hybridization and Quantitative Reverse Transcription Polymerase Chain Reaction Performed by Central Laboratories

2010 ◽  
Vol 28 (28) ◽  
pp. 4300-4306 ◽  
Author(s):  
Frederick L. Baehner ◽  
Ninah Achacoso ◽  
Tara Maddala ◽  
Steve Shak ◽  
Charles P. Quesenberry ◽  
...  
Keyword(s):  
Growth Factor Receptor ◽  
Oncotype Dx ◽  
Her2 Status ◽  
Rt Pcr ◽  
Her2 Positive ◽  
Polysomy 17 ◽  
Epidermal Growth ◽  
Polymerase Chain ◽  
Control Study

Purpose The optimal method to assess human epidermal growth factor receptor 2 (HER2) status remains highly controversial. Before reporting patient HER2 results, American Society of Clinical Oncology (ASCO)/College of American Pathologists (CAP) guidelines mandate that laboratories demonstrate ≥ 95% concordance to another approved laboratory or methodology. Here, we compare central laboratory HER2 assessed by fluorescence in situ hybridization (FISH) and quantitative reverse transcriptase polymerase chain reaction (RT-PCR) using Oncotype DX in lymph node–negative, chemotherapy-untreated patients from a large Kaiser Permanente case-control study. Patients and Methods Breast cancer specimens from the Kaiser–Genomic Health study were examined. Central FISH assessment of HER2 amplification and polysomy 17 was conducted by PhenoPath Laboratories (ratios > 2.2, 1.8 to 2.2, and < 1.8 define HER2 positive, HER2 equivocal, and HER2 negative, respectively). HER2 expression by RT-PCR was conducted using Oncotype DX by Genomic Health (normalized expression units ≥ 11.5, 10.7 to < 11.5, and < 10.7 define HER2 positive, HER2 equivocal, and HER2 negative, respectively). Concordance analyses followed ASCO/CAP guidelines. Results HER2 concordance by central FISH and central RT-PCR was 97% (95% CI, 96% to 99%). Twelve percent (67 of 568 patients) and 11% (60 of 568 patients) of patients were HER2 positive by RT-PCR and FISH, respectively. HER2-positive patients had increased odds of dying from breast cancer compared with HER2-negative patients. Polysomy 17 was demonstrated in 12.5% of all patients and 33% of FISH-positive patients. Nineteen of 20 FISH-positive patients with polysomy 17 were also RT-PCR HER2 positive. Although not statistically significantly different, HER2-positive/polysomy 17 patients tended to have the worst prognosis, followed by HER2-positive/eusomic, HER2-negative/polysomy 17, and HER2-negative/eusomic patients. Conclusion There is a high degree of concordance between central FISH and quantitative RT-PCR using Oncotype DX for HER2 status, and the assay warrants additional study in a trastuzumab-treated population.

Multigene profiling to identify clinically relevant actionable mutations in head and neck cancers: An Indian study.

2020 ◽  
Vol 38 (15_suppl) ◽  
pp. e18529-e18529
Author(s):  
Sateesh S. Kunigal ◽  
Shalini Thakur ◽  
Yogesh Shivkumar ◽  
Sheela M L ◽  
Krishna CR ◽  
...  
Keyword(s):  
Head And Neck ◽  
Treatment Outcomes ◽  
Point Mutations ◽  
Progression Free Survival ◽  
Poor Response ◽  
Standard Of Care ◽  
Response To Therapy ◽  
Treatment Modalities ◽  
Prognostic Indicators ◽  
Response To Chemotherapy

e18529 Background: Head and Neck squamous cell carcinoma (HNSCC) represents approximately 5-10% of malignancies worldwide. Despite the advances in treatment modalities there lies a disparity in response, owing to the underlying molecular and genetic make up. In the era of precision medicine, it is important to profile each disease and customise treatment to improve the outcomes by identifying predictive or prognostic indicators at the molecular and genetic level. Methods: 100 patients with HNSCC, diagnosed at our centre from April 2015-17 were profiled by targeted deep sequencing for hotspot mutations in 48 cancer-related genes using Illumina’s TSCAP panel and MiSeq technology. The average coverage across 220 hot spots was greater than 1000X. Data was processed using Strand Avadis NGS. Mutations identified in the tumor were assessed for ‘actionability’ i.e. response to therapy and impact on prognosis. The clinical data for these patients was collected retrospectively and analysed for treatment outcomes with standard therapy. The association of mutations with habits, stage at presentation, poor treatment outcomes and prognosis was also analysed. Results: Somatic variants were detected in 63.1% of cases. Genetic aberrations were identified in major RAS/RAF signalling pathway in 10 % of cases out of which HRAS activating mutations were the most common (n=4). HRAS was co-mutated with PI3KCA (n=3) and PTEN deletions (n=2). Based on the result, Cetuximab was discontinued in 2 patients with metastatic HNSCC. Other targetable mutations included ATM (n=1), STK11 (n=1), RB1 (n=1) and BRAF (n=1). Disruptive and non-disruptive mutations in TP53 alone were found in 43.8% of H&N cancers, varying widely among different histology with variable treatment outcome. Interestingly all metastatic/recurrent patients, with very short progression free survival of 9-12 months (PFS) post Cisplatinum based chemotherapy, were found to have mutated TP53. TP53 was also found to be co- mutated with ATM gene (n=1), an important prognostic marker, indicating poor response to chemotherapy and radiotherapy. The patients with recurrence and poorer prognosis (n =5) showed point mutations in codon regions 173 -273. Conclusions: Indian patients with HNSCC display a wide mutational landscape with specific actionable targets. Although traditional treatment modalities remain the standard of care, specific mutations may be utilised to customise treatment, prognosticate and predict outcomes.

scholarly journals Detection of ALL-1/AF4 fusion transcript by reverse transcription- polymerase chain reaction for diagnosis and monitoring of acute leukemias with the t(4;11) translocation

Blood ◽  
1993 ◽  
Vol 82 (10) ◽  
pp. 2943-2947 ◽  
Author(s):  
A Biondi ◽  
A Rambaldi ◽  
V Rossi ◽  
L Elia ◽  
C Caslini ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Fusion Transcript ◽  
Response To Therapy ◽  
Molecular Evidence ◽  
Chromosome 11 ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Acute Leukemias ◽  
Polymerase Chain

Abstract The chromosomal breakpoints of t(4;11) translocation of acute lymphoblastic leukemia (ALL) have been recently identified at molecular level and shown to involve the AF4 (FEL) gene on chromosome 4 and the ALL-1 (MLL, Hrx) gene on chromosome 11. The ALL-1/AF4 fusion gene is transcribed into a chimeric mRNA. Using primer sets derived from ALL-1 and AF4 cDNAs by reverse transcription-polymerase chain reaction (RT- PCR), we were able to amplify the breakpoint sites of the fusion transcript of all 15 ALL cases with karyotypic or molecular evidence of the t(4;11). DNA fragments of different size were obtained as the consequence of different breakpoints on chromosome 11 and the presence of alternative splicing of ALL-1 exon 8. The feasibility of monitoring the residual cells carrying the t(4;11) in 2 ALL patients with different clinical outcome was evaluated. Overall, the presented results provide evidence that RT-PCR can be used as a rapid method for detecting this chromosomal abnormality and following the patient's response to therapy.

scholarly journals Detection of ALL-1/AF4 fusion transcript by reverse transcription- polymerase chain reaction for diagnosis and monitoring of acute leukemias with the t(4;11) translocation

Blood ◽  
1993 ◽  
Vol 82 (10) ◽  
pp. 2943-2947 ◽  
Author(s):  
A Biondi ◽  
A Rambaldi ◽  
V Rossi ◽  
L Elia ◽  
C Caslini ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Fusion Transcript ◽  
Response To Therapy ◽  
Molecular Evidence ◽  
Chromosome 11 ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Acute Leukemias ◽  
Polymerase Chain

The chromosomal breakpoints of t(4;11) translocation of acute lymphoblastic leukemia (ALL) have been recently identified at molecular level and shown to involve the AF4 (FEL) gene on chromosome 4 and the ALL-1 (MLL, Hrx) gene on chromosome 11. The ALL-1/AF4 fusion gene is transcribed into a chimeric mRNA. Using primer sets derived from ALL-1 and AF4 cDNAs by reverse transcription-polymerase chain reaction (RT- PCR), we were able to amplify the breakpoint sites of the fusion transcript of all 15 ALL cases with karyotypic or molecular evidence of the t(4;11). DNA fragments of different size were obtained as the consequence of different breakpoints on chromosome 11 and the presence of alternative splicing of ALL-1 exon 8. The feasibility of monitoring the residual cells carrying the t(4;11) in 2 ALL patients with different clinical outcome was evaluated. Overall, the presented results provide evidence that RT-PCR can be used as a rapid method for detecting this chromosomal abnormality and following the patient's response to therapy.

scholarly journals The effect of chemo- and chemoimmunotherapy on the presence of circulating melanoma cells in peripheral blood. Preliminary results.

1998 ◽  
Vol 45 (1) ◽  
pp. 95-102 ◽  
Author(s):  
J Szenajch ◽  
A Kozak ◽  
A A Kruszewski ◽  
E Babiej ◽  
M Chomicka ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Advanced Disease ◽  
Residual Disease ◽  
Melanoma Cells ◽  
Response To Therapy ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
The One ◽  
Therapy Protocol

Reverse transcription and polymerase chain reaction (RT/PCR) with primers specific for tyrosinase allow for a new method of early detection of individual melanoma cells in peripheral blood. Using this test the effect of chemo- and chemoimmunotherapy on the spread of early micrometastatic cancer cells has been evaluated. No significant correlations have been found between RT/PCR results on the one hand and stage of disease, a kind of the therapy protocol used and usage of the therapy as an adjuvant or palliative on the other hand. Thus, although the RT/PCR test for detection of circulating individual melanoma cells might help in identification of minimal residual disease in some patients, it has no application for routine staging of more advanced disease and in monitoring the response to therapy.

Prognostic Significance of VLA4 and CD44 Expression by Flow Cytometry in Multiple Myeloma (MM)

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 4975-4975
Author(s):  
Wasif Riaz ◽  
John Pham ◽  
Ling Zhang ◽  
Lynn Moscinski ◽  
Lori A Hazlehurst ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Flow Cytometry ◽  
Prognostic Significance ◽  
Response To Therapy ◽  
Age At Diagnosis ◽  
Poor Survival ◽  
First Line ◽  
Cd44 Expression ◽  
First Line Therapy ◽  
Line Therapy

Abstract Abstract 4975 Background: Our group has developed a peptide referred to as HYD1 which binds VLA-4 integrin/CD44 containing complex and induces necrotic cell death in myeloma cells (Nair et al Mol Cancer Ther 2009, Emmons et al, Mol Cancer Ther, 2011). Furthermore, we previously demonstrated in a small subset of MM patients that HYD1 was more active in specimens obtained from relapsed/refractory patients, a finding that correlated with VLA-4 expression. The frequency of expression and prognostic significance of two key surface markers VLA4 (very late antigen 4) and CD44 in patients with MM remains poorly defined. Methods: We retrospectively reviewed records of patients with MM who had a complete flow cytometry panel (which includes assessment for CD44 and VLA4) at Moffitt Cancer Center between 1/1/2004 and 12/31/2009. CD44 and VLA4 were gated by CD138 expression and we categorized expression as negative, dim, moderate or bright. We collected demographic information, disease related characteristics (including ISS stage, cytogenetics, and baseline laboratory testing) and treatment and outcomes related data (prior therapies, response to first line therapy, follow up and vital status). Responses were per IMWG criteria and for the purpose of this study, a partial remission or better qualified as a response to therapy. High risk cytogenetics (HRC) was defined by the presence of one of the following; 17p deletion, t(4;14), t(14;16) and 13q deletion by metaphase cytogenetics. The study was approved by the IRB at the University of South Florida. Results: A four color flow cytometry panel was available for review on 101 myeloma patients including 57 males, median age at diagnosis was 60 (range 39–83) years. The percentage of ISS stage I, II and III at diagnosis was 36. 4%, 34. 8% and 28. 8% respectively and 28. 7% of the patients had HRC. At the time of the flow panel, 32 patients (31. 7%) were untreated and overall patients had a median of 3 prior therapies (range 0–11). 52 patients (51. 5%), 37 (36. 6%) and 12 (11. 9%) had no, dim or moderate CD44 expression respectively while 26 (26. 3%), 68 (68. 7%) and 5 (5. 1%) had no, dim or moderate VLA4 expression respectively. Subsequently both were sub-grouped according to presence (dim or greater) or absence of expression. A correlation between the expression of CD44 and VLA4 was noted (Fisher's exact p < 0. 001), 45 patients express both markers, 24 express neither markers, 28 express VLA4 but not CD44 while only 4 express CD44 and not VLA4 (p<0. 001). However neither CD44 nor VLA4 expression correlated with prior therapy (untreated versus previously treated), ISS stage, or poor risk cytogenetics (not shown). 76 patients (75. 2%) responded to first line therapy of which 50 expressed VLA4 (66%) while 25 patients did not respond to first line therapy, of which 23 expressed VLA4 (92%) suggesting VLA4 expression is a negative prognostic factor for response to therapy (p=0. 011). Alternatively CD44 did not correlate with response to first line therapy (not shown). The median OS for patients with VLA4 expression was 60. 5 months (95% CI 44. 9–74. 6) while it was 106. 8 months (95% CI 56. 3-Not reached) for patients who did not express VLA4 (log rank p=0. 023). For CD44 expressing patients, the median OS was 65. 8 months (95% CI 49. 3–86. 8) versus 98. 1 months (95% CI 48. 2–106. 8) for patients without CD44 expression (log rank p= 0. 817). On multivariable analysis, age at diagnosis (p=0. 02) and DS stage (p=0. 003) were the only predictors of poor survival while VLA4 expression (p=0. 09) had a trend towards poor survival. Conclusions: While total CD44 expression was not prognostic in this study, the expression of VLA4 was associated with a decreased response to therapy and decreased survival in multiple myeloma patients and validates this marker as a therapeutic target in myeloma. Although total CD44 expression did not correlate with prognosis in this study, expression of specific CD44 splice variants could be prognostic and this does not dismiss CD44 as a target for the novel therapeutic HYD-1 peptide. These studies are currently being pursued by our laboratory. Disclosures: Hazlehurst: Modulation Therapeutics Inc: Patents & Royalties, President and Co-founder Other. Emmons:Modulation Therapeutic Inc: Patents & Royalties.

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