First Evidence for High Incidence of Complete and Sustained Molecular Remissions and Maintenance of Immune Responses in Patients Receiving Consolidation with Y90 Ibritumomab Tiuxetan (90Y-RIT) Post R-CHOP for Newly Diagnosed Advanced Stage High and Intermediate Risk Follicular Lymphoma

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 2701-2701
Author(s):  
Nancy M Pennell ◽  
Matthew Cheung ◽  
Lisa K Hicks ◽  
Eugenia Piliotis ◽  
Kevin R Imrie ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Immune Responses ◽  
Follicular Lymphoma ◽  
B Cell ◽  
Complete Response ◽  
Advanced Stage ◽  
First Line ◽  
Molecular Remission ◽  
Cell Counts

Abstract Abstract 2701 Introduction: The First-Line Indolent Trial (FIT) demonstrated that administration of a single infusion of 90Y-RIT to follicular lymphoma grade 1 or 2 patients with CR or PR after induction chemotherapy could significantly prolong remissions over no further therapy. Only 13% of patients in that trial received rituximab with their first-line chemotherapy and maintenance rituximab was not administered [JCO 2008;26(32):5156–63]. We report a phase II study of safety and efficacy of 90Y-RIT following R-CHOP chemotherapy in patients with high-risk advanced stage FL. Maintenance rituximab is given every 3 months post 90Y-RIT, for 24 months. The protocol accrues up to 33 evaluable patients. First-time novel insights into the biologic and immunologic effects of this combination regimen are presented. The primary endpoint of our study is the final complete response (CR) rate, defined according to the Cheson criteria and measured 3 months after day 1 of the 90Y-RIT therapy. The CR rate among similar risk advanced stage FL patients after R-CHOP is known to be about 20% [Blood 2005;106(12):3725–32]. The current study is designed to detect a significantly greater response rate of 40% with a p value of 0.05 and a power of 80%. Secondary outcomes included molecular remission and immunologic effects. Since molecular remission is associated with prolonged survival [Blood 105(9): 3428–3433], we analyzed quantitative PCR for up to two years post 90Y-RIT therapy on a subset of patients with baseline PCR detectable t(14:18) lymphoma. Methods: In patients who had PCR detectable t(14;18), quantitative real-time PCR was performed on blood and available bone marrow, at baseline, post 6xR-CHOP and Q 3 months post 90Y-RIT treatment for 24 months. The sensitivity of our PCR assay was 1 in 105 cells. Flow cytometry for % B cell clonality was performed at the same time points. T and B cell counts, Ig levels and vaccine serology have been recorded pre and post treatment. Genotyping of FCGR3A polymorphism was performed. Results: 24 pts have been enrolled with a median age of 54 yrs, 79% stage 4, 63% high FLIPI, 38%, intermediate FLIPI. 10 of 19 [53%] patients had a complete response (CR/CRu) to R-CHOP and the remaining 9 showed partial response (PR). One patient died due to sepsis prior to 90Y-RIT. 5 Patients with a partial response post R-CHOP converted to CRu post 90Y-RIT resulting in 14 of 18 (77%) patients who achieved CR/Cru. One patient progressed 6 m post 90Y-RIT, another relapsed at 12 months. One pt developed a secondary malignancy by 9 m. There were no SAEs attributable to the 90Y-RIT treatment. 16 of 24 (67%) had PCR detectable t(14:18) translocations. Quantitative PCR measurements were concordant with flow cytometry. 18 have been evaluated post 90Y-RIT. 12/13 (92%) of these pts became PCR negative in blood. Post 90Y-RIT, one pt relapsed and showed increase in PCR levels. All remaining pts with PCR markers are PCR negative in blood as far as 24 m post 90Y-RIT. CD3+T cell counts remained normal, but CD19+B cells fell below the 1% detection level by flow cytometry during the two yrs of maintenance therapy post 90Y-RIT. 16 patients have been followed for more than 6 m and 8 pts for 18–24 m with no significant increases in the CD19+ B cell numbers. Interestingly, mean IgG levels remained close to normal, but mean IgM levels fell below normal. Memory immune responses to measles and mumps were maintained post chemo-radiotherapy. Antibody titres to Rubella did not change significantly post 90Y-RIT. Conclusions: We found effective eradication of follicular lymphoma from the blood and bone marrow of the high risk lymphoma pts with first line treatment of 6xRCHOP and all but one of our evaluable patients (92 %) achieved molecular remission in blood post 90Y-RIT. Molecular remission was sustained with maintenance rituximab after 90Y-RIT up to 2 years. Longer follow-up will be necessary to determine the precise response duration. Our molecular response results compare favourably with those of the FIT trial. [JCO 2009;27(34):6094–00]. In our trial, IgG levels remain close to normal indicating that memory B cells are intact and this was consistent with no significant change in titres to common previously vaccinated pathogens such as rubella. A reduction in IgM levels indicating potential functional consequences in mounting primary humoral responses was documented. Disclosures: Off Label Use: Zevalin in first line follicular lymphoma.

Sustained Immune Competency and Long Term Molecular Remissions in FL Patients with FLIPI Risk Factors >1, Treated Front Line with R-CHOP Followed by Consolidative 90 Y-Radioimmunotherapy and Maintenance Rituximab

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 3681-3681
Author(s):  
Neil L Berinstein ◽  
Nancy M Pennell ◽  
Mary-Anne Cussen ◽  
Kevin R Imrie ◽  
Eugenia Piliotis ◽  
...  
Keyword(s):  
Risk Factors ◽  
Flow Cytometry ◽  
Follicular Lymphoma ◽  
Complete Response ◽  
Line Treatment ◽  
First Line ◽  
Molecular Remission ◽  
Cell Counts ◽  
First Line Treatment

Abstract Abstract 3681 Introduction: The First-Line Indolent Trial (FIT) demonstrated that administration of a single infusion of 90Y-RIT to follicular lymphoma grade 1 or 2 patients with CR or PR after induction chemotherapy could significantly prolong time to progression versus no further therapy. [JCO 2008;26(32):5156–63]. Recently, Fowler et al (ASH 2011, abstract 99) have reported on a chemoimmunotherapy approach followed by both RIT consolidation and rituximab maintenance for advanced stage FL patients with FLIPI risk factors >2. However, molecular status (PCR for t[14;18]) was not reported beyond one year of follow-up. We report a phase II study of safety and efficacy of 90Y-RIT following R-CHOP chemotherapy in patients with FLIPI=2–5, advanced stage FL. Maintenance rituximab is given every 3 months (m) post 90Y-RIT, for 24 m. Planned accrual is 33 evaluable patients with extensive two year molecular and immunologic follow-up. Novel insights into the biologic and immunologic effects of this combination regimen are presented. The primary endpoint of our study is the final complete response (CR) rate, defined according to the Cheson criteria and measured 3 m after day 1 of the 90Y-RIT therapy. Secondary outcomes included molecular remission and immunologic effects. Methods: In patients who had PCR detectable t[14;18] in baseline diagnostic specimens, quantitative real-time PCR was performed on blood and available bone marrow, at baseline, post 6xR-CHOP and Q 3 months post 90Y-RIT treatment for 24 m. The sensitivity of our PCR assay was 1 in 105cells. Flow cytometry for % B cell clonality was performed at the same time points. T and B cell counts, Ig levels and vaccine serology have been recorded pre and post treatment. Results: We have enrolled 26 patients with a median age of 54 yrs, 80% stage 4, 58%, intermediate FLIPI=2, 42% high FLIPI=3–5. Sixteen of 26 [62%] patients had a complete response (CR/CRu) to R-CHOP and the remaining 10 [40%] showed partial response (PR). One patient died due to sepsis prior to 90Y-RIT. Five patients with a partial response post R-CHOP converted to CRu post 90Y-RIT. A total of 19 of 24 (79%) patients who received 90Y-RIT, achieved CR/Cru. Post 90Y-RIT, three patients have relapsed. One other developed a secondary malignancy by 9m. The treatment has been most favourable for patients with FLIPI=2, where 13/14 (93%) remain progression-free [median follow-up= 32 m, range= 12.5–62 m]. There were no SAEs attributable to the 90Y-RIT treatment. Seventeen patients had PCR detectable t(14:18) translocations. Quantitative PCR measurements were concordant with flow cytometry. Of these, 16, were evaluated post 90Y-RIT and 15/16 (94%) of these patients became PCR negative in blood. Post 90Y-RIT, 2 patients showed increase in PCR levels and relapsed clinically. All remaining pts with PCR markers are PCR negative in blood as far as 24 m post 90Y-RIT. CD3+T cell counts remained normal, but CD19+B cells fell below the 1% detection level by flow cytometry during the two yrs of maintenance therapy post 90Y-RIT. Interestingly, mean IgG levels remained close to normal, but mean IgM levels fell below normal. Memory immune responses to measles and mumps were maintained post chemo-radiotherapy. Antibody titres to Rubella did not change significantly post 90Y-RIT. No HAMA response has been detected in any of the patients. Conclusions: We found effective eradication of follicular lymphoma from the blood and bone marrow of the high risk lymphoma pts with 2 or more FLIPI risk factors with first line treatment of 6xRCHOP and all but one of our evaluable patients (94%) achieved molecular remission in blood post 90Y-RIT. Molecular remission was sustained after 90Y-RIT up to 2 years, considerably longer than that reported by Fowler et al (ASH 2011, abstract 99). Longer follow-up with annual monitoring is planned to determine the precise response duration. The progression-free survival rates are similar or more favourable to previous reports [Blood 2006;108:1504–1508, J Clin Exp Hem. 2012; 52,no. 1]. IgG levels remain close to normal indicating that memory B cells are intact and this was consistent with no significant change in titres to common previously vaccinated pathogens such as rubella. The significance of persistent reductions on IgM levels is unclear. Acknowledgments: This study was sponsored by Bayer Canada and Spectrum Pharmaceuticals. Disclosures: Berinstein: Spectrum Pharma: Clinical Advisory Board Other. Off Label Use: Zevalin for first line treatment of aggressive follicular lymphoma.

scholarly journals The Prognostic Value of IgH Clonality in Bone Marrow with Negative Pathology in B Cell Lymphoma

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 5298-5298
Author(s):  
Kazuhiro Toyama ◽  
Ayato Tsukamoto ◽  
Sho Yamazaki ◽  
Fumihiko Nakamura ◽  
Mineo Kurokawa
Keyword(s):  
Bone Marrow ◽  
Prognostic Factor ◽  
B Cell ◽  
Research Funding ◽  
Bone Marrow Involvement ◽  
Pharmaceutical Research ◽  
Complete Response ◽  
Pathological Examination ◽  
First Line ◽  
First Line Therapy

Abstract Background Bone marrow infiltration is widely accepted as an important prognostic factor in malignant lymphomas. Although multiplex polymerase chain reaction for the detection of clonal immunoglobulin heavy chain (IgH) gene rearrangements is a helpful tool for the confirmation of minimal infiltration in B cell lymphomas, it has not been elucidated that IgH clonality of the bone marrow at the initial evaluation is a valuable prognostic factor and a predicting marker for the initial therapy in the patients with B cell lymphomas who had a bone marrow biopsy with negative pathology. Methods All patients with DLBCL (n=61) and indolent B cell non-Hodgkin's lymphomas (iB-NHL) including FL, MCL and MALT lymphoma (n=51) who had a bone marrow biopsy and a bone marrow IgH clonality obtained at diagnosis in our institute it the period between March 2002 and 2016 were included in this study. The clinical information including the overall survival (OS), progression free survival (PFS), laboratory findings, backgrounds, and the response to the first line therapy were collected from the clinical charts. Result Bone marrow infiltration by pathological examination was reported in 20% (12/61) of DLBCL patients and in 24% (12/51) of iB-NHL patients (Path-pos group). In the patients with negative pathological report, 21% of the DLBCL patients (13/61) and 33% of the iB-NHL patients (17/51) were positive bone marrow involvement by PCR (Path-neg/PCR-pos group). The other 59% of the DLBCL patients (36/61) and 43% of the iB-NHL patients (22/51) were negative bone marrow involvement by both pathological examination and PCR (Path-neg/PCR-neg group). In DLBCL cohort, the 5-year OS rates in Path-pos, Path-neg/ PCR-pos, and Path-neg/ PCR-neg groups were 46.9% (95% CI, 13.2% to 75.3%), 92.3% (95% CI, 56.6% to 98.9%), and 85.5% (95% CI, 68.7% to 93.7%), respectively (p=0.0735). The 5-year PFS rates in Path-pos, Path-neg/PCR-pos, and Path-neg/PCR-neg group were 33.3% (95% CI, 10.3% to 58.8%), 76.9% (95% CI, 44.2% to 91.9%), and 71.1% (95% CI, 52.9% to 83.3%), respectively (p=0.0331). There were no significant difference between Path-neg/ PCR-pos group and Path-neg/ PCR-neg group in the 5-year OS and PFS rates (p=0.698). In DLBCL patients, the positivity of pathological examination of bone marrow was identified as a valuable prognostic factor, but the positivity of PCR was not a prognostic factor. In iB-NHL cohort, the 5-year OS rates in Path-pos, Path-neg/PCR-pos, and Path-neg/PCR-neg groups were 77.5% (95% CI, 44.8% to 92.3%), 100% (95% CI, 100% to 100%), and 94.4% (95% CI, 66.6% to 99.2%), respectively (p=0.114). The 5-year PFS rates in Path-pos, Path-neg/PCR-pos, and Path-neg/PCR-neg group were 45.7% (95% CI, 17.3% to 70.5%), 46.8% (95% CI, 19.6% to 70.2%), and 87.1% (95% CI, 57.3% to 96.6%), respectively (p=0.152). There were almost no difference between Path-pos, Path-neg/PCR-pos, and Path-neg/PCR-neg group in the 5-year OS rate, but the 5-year PFS rate in Path-pos and Path-neg/PCR-pos group tend to be lower than in Path-neg/PCR-neg group (p=0.0641 and p=0.0891, respectively). In iB-NHL cohort, in addition to the positivity of pathological examination, the positivity of PCR is possible to be a prognostic factor. Univariate analysis of PFS in pathology negative patients in iB-NHL revealed that performance status was a significant prognostic factor, and IgH clonality also seemed to be a prognostic factor. Multivariate analysis of PFS in pathology negative patients in iB-NHL showed that there was no significant independent risk factor. In the response to the first line therapy in DLBCL cohort, the rate of complete response in Path-pos, Path-neg/PCR-pos, and Path-neg/PCR-neg groups were 83.3%, 90.9%, and 80.0%, respectively (p=0.8871). In iB-NHL cohort, the rate of complete response were 60.0%, 71.4%, and 82.4%, respectively (p=0.4623). In each cohort, there were no difference between each groups in overall response rate and complete response rate. Conclusion Positive IgH clonality with negative pathology in bone marrow is a valuable prognostic factor for PFS at diagnosis in the patients with iB-NHL. Disclosures Kurokawa: Eizai: Research Funding; MSD: Honoraria, Research Funding; Chugai Pharmaceutical: Research Funding; Teijin Pharma: Research Funding; Takeda Pharmaceutical: Research Funding; Pfizer: Research Funding; Ono Pharmaceutical: Honoraria, Research Funding; Astellas Pharma: Research Funding; Sumitomo Dainippon Pharma: Research Funding; Nippon Sinyaku: Honoraria, Research Funding; Otsuka Pharmaceutical: Research Funding; Kyowa Hakko Kirin: Honoraria, Research Funding.

Clinical Translation of a Prognostic Follicular Lymphoma Signaling Profile

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 636-636
Author(s):  
Jonathan M Irish ◽  
Debra K Czerwinski ◽  
Garry P. Nolan ◽  
Ronald Levy
Keyword(s):  
Flow Cytometry ◽  
Follicular Lymphoma ◽  
B Cell ◽  
Tumor Cells ◽  
Cell Counting ◽  
Color Flow ◽  
Individual Measurement ◽  
Risk Of Death ◽  
Cell Counts ◽  
Bcr Signaling

Abstract Abstract 636 Introduction: Phospho-flow cytometry signaling profiles have great potential to identify and monitor negative prognostic subpopulations of cells within tumors. We recently reported that altered B cell receptor (BCR) signaling identifies a subset of lymphoma negative prognostic (LNP) cells within follicular lymphoma (FL) tumors from patients with shorter overall survival (Irish et al., PNAS 2010). Each 1% increase in LNP cells observed within the tumor at diagnosis increased the patient's annual risk of death by 2.5%. However, before signaling profiles such as this can be used in the clinic, the assay must be simplified and translated from the complex and time-consuming version used in research studies. Here we report a prognostic signaling profile suitable for follicular lymphoma clinical specimens of less than one million cells. Methods: To simplify LNP cell counting in FL patient specimens, detection of six key phospho-protein readouts for BCR signaling was combined on one cytometer channel, creating a single readout for BCR network signaling activity (termed p-BCR network). This combined stain measured phosphorylation of SYK, PLCγ, BLNK, Src family kinases (SFK), STAT5, and ERK using Alexa 488 dye conjugates of each phospho-specific antibody. LNP cells are defined by a lack of BCR signaling response at all of these phospho-protein readouts. Upon receipt of a patient's tumor sample, 0.25 million cells were stimulated for 4 minutes by polyclonal F(ab')2 against IgM and IgG (10 μ g/mL each; α-BCR), left unstimulated as a negative control, or exposed to a positive control stimulus (α-BCR + H2O2). Samples were stained with a four-color flow cytometry panel detecting p-BCR network, lymphoma B cell markers BCL2 and CD20, and CD3 or CD5 as a marker for T cells. Approximately 50,000 lymphoma B cells were collected and LNP cells quantified as before. The p-BCR network assay typically required 2h to complete. In parallel, LNP cells were quantified in samples from the same specimens using the original method, where phospho-proteins were detected individually using a set of staining panels. An additional control staining panel measured total tyrosine phosphorylation (p-Y) in place of p-BCR network staining in the four-color panel. Results: LNP cell counts measured by combined p-BCR network stain using 0.75 million FL tumor cells total were equivalent to those obtained by individual measurement of phospho-proteins (R2 = 0.98). Critically, in the combined p-BCR network stain the same patterns of signaling in lymphoma cell subpopulations were observed as when individual phospho-proteins were measured. Thus, not only was the same quantity of LNP cells detected by the p-BCR network stain, the same subpopulations of cells were identified by the two techniques. Total phospho-tyrosine following BCR stimulation was examined as a potential surrogate for p-BCR network staining, but high basal p-Y signaling and additional BCR mediated p-Y signaling obscured the LNP cell subset in some patient specimens. Thus, a total p-Y readout could not be used in place of the p-BCR network readout or detection of individual phospho-protein readouts. Conclusions: A clinical prognostic follicular lymphoma signaling profile can be measured in small specimens of live tumor cells, such as fine needle aspirates, and collected using commonly available flow cytometers. These results create new opportunities to use signaling profiles to identify FL patients who might benefit from clinical trials and to monitor emergence of negative prognostic lymphoma cells over time following therapy. Disclosures: No relevant conflicts of interest to declare.

Follicular Lymphoma in Staging Bone Marrow Specimens: Correlation of Histologic Findings With the Results of Flow Cytometry Immunophenotypic Analysis

2007 ◽  
Vol 131 (2) ◽  
pp. 282-287
Author(s):  
Dan Iancu ◽  
Suyang Hao ◽  
Pei Lin ◽  
S. Keith Anderson ◽  
Jeffrey L. Jorgensen ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Follicular Lymphoma ◽  
B Cell ◽  
Bone Marrow Biopsy ◽  
Cell Population ◽  
Color Flow ◽  
Flow Cytometric Immunophenotyping ◽  
Histologic Evidence ◽  
Flow Cytometric ◽  
Biopsy Specimens

Abstract Context.—Bone marrow (BM) examination is part of the staging workup of lymphoma patients. Few studies have compared BM histologic findings with results of flow cytometric immunophenotyping analysis in follicular lymphoma (FL) patients. Objective.—To correlate histologic findings with immunophenotypic data in staging BM biopsy and aspiration specimens of FL patients. Design.—Bone marrow biopsy specimens of untreated FL patients were reviewed. Histologic findings were correlated with 3-color flow cytometric immunophenotyping results on corresponding BM aspirates. Results.—Bone marrow biopsy specimens (with or without aspirates) of 114 patients with histologic evidence of FL in BM were reviewed. There were 76 bilateral and 38 unilateral biopsies performed, resulting in 190 specimens: 187 involved by FL and 3 negative (in patients with a positive contralateral specimen). The extent of BM involvement was <5% in 32 (17.1%), ≥5% and ≤25% in 102 (54.6%), >25% and ≤50% in 27 (14.4%), and >50% in 26 (13.9%) specimens. The pattern of involvement was purely paratrabecular in 81 (43.3%), mixed in 80 (42.8%), and purely nonparatrabecular in 26 (13.9%). Immunophenotyping was only performed unilaterally, on BM aspirates of 92 patients, and was positive for a monoclonal B-cell population in 53 (57.6%) patients. Immunophenotyping was more often negative when biopsy specimens showed FL with a purely paratrabecular pattern. For comparison, we assessed 163 FL patients without histologic evidence of FL in BM also analyzed by flow cytometric immunophenotyping. A monoclonal B-cell population was identified in 5 patients (3%). Conclusions.—Our data suggest that 3-color flow cytometric immunophenotyping adds little information to the evaluation of staging BM specimens of FL patients.

Unrelated Donor Marrow Transplantation for B-Cell Chronic Lymphocytic Leukemia After Using Myeloablative Conditioning: Results From the Center for International Blood and Marrow Transplant Research

2005 ◽  
Vol 23 (24) ◽  
pp. 5788-5794 ◽  
Author(s):  
Steven Z. Pavletic ◽  
Issa F. Khouri ◽  
Michael Haagenson ◽  
Roberta J. King ◽  
Philip J. Bierman ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Chronic Lymphocytic Leukemia ◽  
B Cell ◽  
Marrow Transplantation ◽  
Chronic Gvhd ◽  
Complete Response ◽  
Lymphocytic Leukemia ◽  
Unrelated Donor ◽  
Myeloablative Conditioning ◽  
Cell Chronic Lymphocytic Leukemia

Purpose To determine the role of myeloablative conditioning and unrelated donor (URD) bone marrow transplantation in the treatment of patients with advanced B-cell chronic lymphocytic leukemia (CLL). Patients and Methods A total of 38 CLL patients received a matched URD transplant using bone marrow procured by the National Marrow Donor Program. The median age was 45 years (range, 26 to 57 years), the median time from diagnosis was 51 months, and the median number of prior chemotherapy regimens was three. Fifty-five percent of patients were chemotherapy refractory and 89% had received fludarabine. Conditioning included total-body irradiation in 92% of patients. Graft-versus-host disease (GVHD) prophylaxis consisted of methotrexate with cyclosporine or tacrolimus for 82% of patients. Results Twenty-one patients (58%) achieved complete response and six (17%) achieved partial response. Incidences of grades 2 to 4 acute GVHD were 45% at 100 days and incidences of chronic GVHD were 85% at 5 years. Eleven patients are alive and disease free at a median of 6 years (range, 3.0 to 9.0 years). Five-year overall survival, failure-free survival, disease progression rates, and treatment-related mortality (TRM) were 33%, 30%, 32%, and 38% respectively. Conclusion These data demonstrate that lasting remissions can be achieved after URD transplantation in patients with advanced CLL. High TRM suggest that myeloablative conditioning and HLA-mismatched donors should be avoided in future protocols, and it is mandatory to investigate transplant strategies with a lower morbidity and mortality, including the use of nonmyeloablative regimens.

scholarly journals Mesenchymal Stromal Cells Orchestrate Follicular Lymphoma Cell Niche Through the CCL2-Dependent Recruitment and Polarization of Monocytes

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 1566-1566
Author(s):  
Fabien Guilloton ◽  
Gersende Caron ◽  
Cédric Ménard ◽  
Céline Pangault ◽  
Patricia Amé-Thomas ◽  
...  
Keyword(s):  
Bone Marrow ◽  
B Cells ◽  
Cell Growth ◽  
Follicular Lymphoma ◽  
B Cell ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Gene Set Enrichment Analysis ◽  
Cell Niche

Abstract Abstract 1566 Accumulating evidence indicates that infiltrating stromal cells contribute directly and indirectly to tumor growth in a wide range of solid cancers and hematological malignancies. In follicular lymphoma (FL), malignant B cells are found admixed with heterogeneous lymphoid-like stromal cells within invaded lymph nodes and bone marrow (BM). In addition, in vitro functional studies have underlined that mesenchymal cells recruit malignant FL B cells and protect them from spontaneous and drug-induced apoptosis. In particular, we have previously demonstrated that mesenchymal stromal cells (MSC) efficiently support in vitro FL B-cell survival, especially after their engagement towards lymphoid differentiation through treatment with TNF-α and Lymphotoxin-α1β2 (TNF/LT) or after coculture with malignant B cells. However, the mechanisms of this supportive activity remain largely unknown. In this study, we used Affymetrix U133 Plus 2.0 microarrays, to compare the gene expression profile (GEP) of bone marrow-derived MSC (BM-MSC) obtained from 10 FL patients at diagnosis versus 6 age-matched healthy donors (HD). In these conditions, neither the CFU-F concentration in the BM nor the cumulative population doubling of BM-MSC significantly differed between HD and FL patients. Unsupervised analysis was able to perfectly segregate FL-MSC from HD-MSC and we identified, using supervised analyzes, a list of 408 probesets defining FL-MSC signature, including 320 nonredundant genes upregulated in FL-MSC compared to HD-MSC. We then defined the GEP of human lymphoid-like stroma using HD-MSC treated in vitro by TNF/LT and demonstrated, by a Gene Set Enrichment Analysis (GSEA) approach, that the FL-MSC signature is significantly enriched for genes associated with a lymphoid-like commitment. Interestingly, CCL2 was strongly overexpressed by FL-MSC, was upregulated in HD-MSC by coculture with malignant B cells, and was detected at a higher level in FL BM plasma compared to normal BM plasma (504.4 pg/mL [23.8-4413] versus 33.9 pg/mL [5-126.1]; P <.01). In agreement, FL-MSC triggered a more potent CCL2-dependent monocyte migration than HD-MSC. Moreover, FL-MSC and macrophages cooperated to sustain malignant B-cell growth through both protection from apoptosis and enhancement of cell proliferation. Finally, FL-MSC promoted monocyte differentiation towards a proangiogenic LPS-unresponsive phenotype close to that of tumor-associated macrophages. We unraveled a key role for the Notch pathway in this process and identified an overexpression of JAGGED1 in FL-MSC compared to HD-MSC. Altogether, these results highlight the complex role of FL stromal cells that promote direct tumor B-cell growth and orchestrate FL cell niche. The identification and characterization of this intricate network of cell interactions may provide novel therapeutic targets in this disease. Disclosures: No relevant conflicts of interest to declare.

scholarly journals High-Dose Chemotherapy with Bendamustin and Melphalan Improves the Rate of Complete Remission in Myeloma Patients in First Remission Compared to Standard Melphalan Alone

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 39-40
Author(s):  
Sarah Farag ◽  
Ulrike Bacher ◽  
Myriam Legros ◽  
Daniel Betticher ◽  
Jean-Marc Lüthi ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Complete Remission ◽  
Median Time ◽  
Maintenance Treatment ◽  
Induction Treatment ◽  
High Dose ◽  
First Line ◽  
Term Survival ◽  
Pulmonary Failure

Introduction: Consolidation of first-line induction treatment in myeloma (MM) patients (pts) with 200 mg/m2 melphalan chemotherapy (HDCT) and autologous stem cell transplantation (ASCT) was established as standard of care three decades ago. However, definite cure in myeloma patients remains exceptional due to residual disease escaping intensive treatment, and almost all patients will ultimately relapse at earlier or later time points following ASCT. Thus, improving efficacy of HDCT in MM remains an unresolved issue. Methods: We performed a phase-II randomized trial comparing standard 200 mg/m2 Melphalan (Mel) HDCT to experimental HDCT treatment with 200 mg/m2 bendamustine, a bifunctional alkylating agent, given at days -4 and -3, combined with 200 mg/m2 melphalan split on days -2 and -1 at 100 mg/m2 (BenMel) before ASCT in MM pts. Patients had up to four cycles of first-line induction treatment with bortezomib, lenalidomide and dexamethasone. After ASCT, pts received lenalidomide maintenance treatment for two years. The primary endpoint was to show a 15% improvement of the rate of complete remission (sCR+CR) after HDCT with BenMel compared to Mel alone. MRD assessment from the bone marrow was performed by multiparameter flow cytometry after hematological engraftment following HDCT/ASCT. MRD negativity was defined as clonal plasma cells below 10(-5). Results: We randomized 120 myeloma pts (60 patients in each arm), with high-risk genetic abnormalities present in 21.3% of the patients. The median age was 63 years (range 35-74). The sCR/CR rate after ASCT before initiation of lenalidomide maintenance treatment was better in the BenMel arm compared to Mel alone (70.0% vs 51.7%; p=.039). The post-ASCT remission rates in detail were sCR 40.0% vs 31.7% (p=.341); CR 30.0% vs 20.0% (p=.205); VGPR 16.7% vs 33.3% (p=.035); and PR 13.3% vs 15.0% (p=.793). MRD negativity assessed in the bone marrow by flow cytometry was observed in 26 (45.6%) of the BenMel treated pts compared to 22 (37.9%) of the Mel pts. Median time until neutrophil engraftment was 11 days after BenMel vs 12 days after Mel (p=.096), and median time until platelet engraftment was 13 days in both arms (p=0.367); all pts had full engraftment of both cell lineages. Prolonged hospitalization duration was seen in BenMel pts (median 19 vs 18 days; p=.006) due to the longer BenMel treatment administration. Fully reversible acute renal insufficiency occurred in three (5%) BenMel pts compared to none of the Mel pts (p=.250). No treatment-related mortality was seen in both groups. ICU admissions were necessary in 3 pts (5%) in the BenMel group (ARDS, septic shock, pulmonary failure), and 2 Mel treated pts (3.3%; due to pulmonary failure and decompensated cardiomyopathy). The PFS rates at 12 months were 95% in BenMel pts and 91% in Mel treated pts (p=.551). OS at 12 months was 96% for both groups (p=.262), and median PFS and OS were not reached in both groups. Conclusions: Our data confirm that high-dose bendamustine combined with melphalan HDCT before ASCT in MM patients is safe and well tolerated. In particular, bendamustine-associated renal toxicity was manageable and reversible in all patients, and hematopoietic engraftment was comparable to standard melphalan HDCT. HDCT with BenMel improves the sCR/CR rate compared to standard melphalan alone. Thus, BenMel HDCT before ASCT warrants further investigation aiming to improve the long-term survival rates of MM patients, eventually combined with new maintenance strategies in the post-transplant period. Figure 1 Disclosures No relevant conflicts of interest to declare.

scholarly journals Expert-independent classification of mature B-cell neoplasms using standardized flow cytometry: a multicentric study

2021 ◽  
Author(s):  
Sebastian Böttcher ◽  
Robby Engelmann ◽  
Georgiana Grigore ◽  
Paula Carolina Fernandez ◽  
Joana Caetano ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Differential Diagnosis ◽  
Follicular Lymphoma ◽  
B Cell ◽  
Diagnostic Algorithm ◽  
Lymphocytic Leukemia ◽  
B Cell Lymphomas ◽  
Multicentric Study ◽  
Predictive Values ◽  
Large B Cell

Reproducible expert-independent flow-cytometric criteria for the differential diagnoses between mature B-cell neoplasms are lacking. We developed an algorithm-driven classification for these lymphomas by flow cytometry and compared it to the WHO gold standard diagnosis. Overall, 662 samples from 662 patients representing nine disease categories were analyzed at 9 laboratories using the previously published EuroFlow 5-tube-8-color B-cell chronic lymphoproliferative disease antibody panel. Expression levels of all 26 markers from the panel were plotted by B-cell entity to construct a univariate, fully standardized diagnostic reference library. For multivariate data analysis we subsequently utilized Canonical Correlation Analysis of 176 training cases to project the multi-dimensional space of all 26 immunophenotypic parameters into 36 two-dimensional plots for each possible pair-wise differential diagnosis. Diagnostic boundaries were fitted according to the distribution of the immunophenotypes of a given differential diagnosis. A diagnostic algorithm based on these projections was developed and subsequently validated using 486 independent cases. Negative predictive values exceeding 92.1% were observed for all disease categories except for follicular lymphoma. Particularly high positive predictive values were returned in chronic lymphocytic leukemia (99.1%), hairy cell leukemia (97.2%), follicular lymphoma (97.2%) and mantle cell lymphoma (95.4%). Burkitt and CD10+ diffuse large B-cell lymphomas were difficult to distinguish by the algorithm. A similar ambiguity was observed between marginal zone, lymphoplasmacytic, and CD10- diffuse large B-cell lymphomas. The specificity of the approach exceeded 98% for all entities. The univariate immunophenotypic library and the multivariate expert-independent diagnostic algorithm might contribute to increased reproducibility of future diagnostics in mature B-cell neoplasms.

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