Evaluation of Angiogenesis and the CD57+ Lymphocytic Population on Bone Marrow Biopsies (BM) in Multiple Myeloma Patients Treated with Thalidomide (Thal)

Abstract Abstract 5095 Introduction – Multiple myeloma (MM) is a plasma cell dyscrasias characterized by the bone destruction, renal insufficiency, anemia and hypercalcemia. Studies suggest that the disease activity and the degree of angiogenesis in the bone marrow (BM) are related. It has been shown that increased microvessel density (MVD) in MM patients`BM specimens is associated with poor prognosis, and BM MVD at diagnosis is an important prognostic factor for survival of patients. Due to the anti-angiogenic effect, Thal is becoming consolidated as a therapeutic option for the treatment of MM. The presence of cytotoxic CD57+ T lymphocytes in the BM in MM patients who have never been treated correlates with the clinical progression of the patients. The present study has the objective of presenting findings of the anti-angiogenic effect of thalidomide, correlating with the reduction of MVD in the obtained response, as well as verifying the presence of CD57+ lymphocytes before and after the treatment, correlating this with the rate response. Materials and methods– BM were collected from the posterior iliac crest in MM patients who had never been treated at least 12 weeks after having initiated treatment with Thal or up to the fourth week after discontinuing its use. Patients who had previously received Thal were excluded. The bone marrow biopsies were done by two pathologists who were blind as to the disease treatment phase. Angiogenesis was estimated based on the MVD, utilizing the anti-CD34 antibody as an immunohistochemical marker. The slides were photographed at 03 sites of densely concentrated vessels selected by counting under 400X magnification. The final density of the microvessel site median was determined. The CD57+ lymphocyte analysis was made for plasmocyte concentration zones, determining the final count using the median of three sites. Statistical analysis was performed with the SPSS 15.0 for Windows, a t-parametric test for equal averages and Spearman correlation. Results– There was a total of 20 patients (pre- and post-treatment). The median age was 64 (40–82 years), 65% of the cases being male. The Durie-Salmon Staging distribution: IIA/B= 10 % and IIIA/B=90%, and ISS: 2=45% and 3=35%. The IgG isotype was present in 70% of the cases. The therapeutic schedule made use of target doses of thalidomide at 200mg/d. Fourteen patients utilized the Thal and dexamethasone (TD) schedule, four patients, cyclophosphamide+TD (CTD), one patient, Melphalan+prednisone+Thal (MPT) and one patient combination TD+CTD. Eleven patients received a 90-day treatment between collections, seven, a 120-day treatment, one, a 150-day treatment and one, a 270-day treatment. The median MVD count of pre-thal CD34 was 11.42, and post-thal, 7.17 (p=0.01). The pre-thal CD57 median was 26.42 and the post-treatment, 21.58 (not significant). There was a negative post-CD34 versus overall response correlation (p=0.04) and a positive CD57+ versus overall response correlation (p=0.05). Pre-CD34 versus International Staging System correlations (p=0.01) were observed. Another unexpected observation was the analysis of the gender as an independent variable, it was observed that the post-treatment CD57+ was significantly different between the sexes (p=0.008). Conclusion– This study confirms the anti-angiogenic effect of Thal in MM patients, with reduced MVD by CD34 analysis, in addition to its correlation with the overall response. It also demonstrates the significant correlation between the post-treatment CD57+ lymphocytic population and the overall response. The unexpected finding was the significant difference in the quantity of lymphocytes present in the post-treatment BM between the genders (increased in men and reduced in women). Furthermore, it was observed that a greater quantity of MVD correlates with the worst ISS staging. Disclosures: No relevant conflicts of interest to declare.

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Anti-Angiogenic Effect of Bortezomib in Multiple Myeloma Patients.

Blood ◽

10.1182/blood.v104.11.4912.4912 ◽

2004 ◽

Vol 104 (11) ◽

pp. 4912-4912

Author(s):

Marianna Politou ◽

Kikeri Naresh ◽

Evangelos Terpos ◽

Danielle Crowley ◽

Irvin Lambert ◽

...

Keyword(s):

Multiple Myeloma ◽

Bone Marrow ◽

Partial Response ◽

Angiogenic Factors ◽

Response To Treatment ◽

Serum Samples ◽

Bone Marrow Biopsies ◽

Significant Difference ◽

Before And After ◽

Angiogenic Effect

Abstract Bortezomib is a proteasome inhibitor, which is an effective treatment for multiple myeloma (MM). Bortezomib inhibits NF-κB and thus enhances apoptosis and leads to reduced levels of growth factors, angiogenic factors and cell adhesion molecules, which are crucial for the growth and survival of myeloma. The aim of this study was to investigate whether bortezomib has an anti-angiogenic effect in MM patients and whether that effect correlates with response to treatment. We have studied the effect of bortezomib on angiogenesis in bone marrow biopsies and serum samples of nine patients with MM, who were treated with bortezomib. The patients studied (6M/3F median age 59 years, range 35–71 years) had received more than 4 lines of treatment before bortezomib administration. Six patients had IgG, one IgA, one non-secretory and one light-chain MM. Bortezomib was given at a dose of 1.3 mg/m2, iv, in 3-week cycles, on days 1, 4, 8, and 11 of each cycle. Microvessel density (MVD) was assessed in bone marrow trephine biopsies before and after 8 cycles of treatment by immunohistochemistry with monoclonal mouse antibodies to CD34 (QBEND-10, DAKO, Denmark). Serum samples were assessed for VEGF and angiogenin levels before and after every cycle of treatment with an ELISA (R&D systems). Five out of 9 patients achieved a partial response (PR), two patients had a minimal response (MR),one achieved a good partial response (GPR) and one a complete response (CR) to bortezomib administration, according to EBMT criteria. In six out of 9 patients there was a decrease of the MVD. More specifically in two of the patients with PR (P2,P4) and the two patients with MR (P5 and P7) there was a significant decrease in the MVD ( 1.7, 3.8, 4.2 and 1.4 fold decrease respectively). Patient 9, who achieved GPR had also a 2.3 fold decrease in MVD. In patients P2,P4,P5 and P6, who received 8 cycles of treatment, further reduction of MVD was noticed with further treatment. In patient 1, who achieved a PR, MVD did not show any significant change after 8 cycles of treatment. The patient relapsed soon after he has completed the treatment. In patient 5, the MVD increased over 2-fold and she relapsed very soon after the 4th cycle and died of disease progression. There was a significant reduction in mean angiogenin levels by cycle 4 (380 ng/ml) when compared with cycle 1 (537ng/ml) (p=0.028). On the contrary, there was no significant difference between the levels of VEGF at cycles 1 and 4 (122.5 pg/ml and 127.2 pg/ml respectively) (p=0.173).All data are shown in Table 1. We conclude that PS-341 may exert its anti-myeloma effect partly through anti- angiogenic mechanisms. Whether Bortezomib acts directly on endothelial cells or indirectly through modulation of the expression of angiogenic factors and angiopoetins which influence endothelial cell proliferartion and survival is unclear. Before treatment After 4th Cycle of treatment After 8th Cycle of treatment response to treatment VEGF(pg/ml)/Ang (ng/ml) MVD vessels/mm 2 VEGF(pg/ml)/Ang (ng/ml) MVD vessels/mm 2 VEGF(pg/ml)/Ang (ng/ml) MVD vessels/mm 2 patient 1 PR 213/642 74.1 813/432 428/361 83.99 Patient 2 PR 172/425 124.7 449/334 77.48 243/371 73.85 patient 3 PR 84/404 61/256 175/2 65 patient 4 PR 160/800 109.55 180/316 37.5 80/359 28.5 patient 5 MR 57/615 195.8 70/517 85.51 94/447 46.1 patient 6 PR 47/459 267 82/335 591.8 patient 7 MR 105.88 75.55 patient 8 GPR 48.89 2 patient 9 CR 135.78 57.14

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The Certainty of a Post-Bone Marrow Diagnosis: A Study of the Yield of Bone Marrow Biopsies in a Community Hospital Setting

Blood ◽

10.1182/blood-2020-142032 ◽

2020 ◽

Vol 136 (Supplement 1) ◽

pp. 34-35

Author(s):

Manasi M. Godbole ◽

Peter A. Kouides

Keyword(s):

Multiple Myeloma ◽

Bone Marrow ◽

Bone Marrow Biopsy ◽

Fever Of Unknown Origin ◽

Conflicts Of Interest ◽

Diagnostic Yield ◽

Hospital Setting ◽

Unknown Origin ◽

Nutritional Deficiencies ◽

Bone Marrow Biopsies

Introduction: Most studies on the diagnostic yield of bone marrow biopsy including the one by Hot et al. have focused on the yield of bone marrow biopsies in diagnosing the source of fever of unknown origin. However, there have not been any studies performed to our knowledge looking at overall practice patterns and yield of bone marrow biopsies for diagnoses other than fever of unknown origin. We aim to determine the most common indications for performing bone marrow biopsies in a community-based teaching hospital as well as the yield of the biopsies in patients with specified and unspecified pre-test indications to estimate the rate of uncertain post-test diagnoses. Methods: We performed a retrospective data collection study at Rochester General Hospital, NY. A comprehensive search was conducted in our electronic medical data to identify all patients who underwent bone marrow biopsies over a 5 year period from January 2011 - December 2016 for indications other than fever of unknown origin. Patient data including demographics, pre-bone marrow biopsy diagnosis and post-bone marrow diagnosis was obtained. All patients above the age of 18 who underwent bone marrow biopsy for indications other than fever of unknown origin or follow up treatment of a hematological malignancy were included. Results: A total of 223 biopsies were performed. The median age was 59 years (age range- 23-95). One hundred and sixteen patients were male and 107 were female. The most common indications for performing bone marrow biopsy were evaluation of the following possible conditions: multiple myeloma (n=54), myelodysplastic syndrome [MDS] (n=47), lymphoma (n=28) and leukemia (n=18) as well as non-specific indications such as pancytopenia (n=40), anemia (n=22) and thrombocytopenia (n=11). The proportion of cases confirmed by bone marrow biopsy was 45/54 (83%) with the pre-marrow diagnosis of multiple myeloma, 34/47 cases (72%) with the pre-marrow diagnosis of MDS, 15/18 (83%) with the pre-marrow diagnosis of leukemia and 13/28 (46%) in those with the pre-marrow diagnosis of rule out lymphoma. Thirteen cases (18%) with possible MDS had post-bone marrow diagnoses of leukemia, anemia of chronic disease, myelofibrosis or medication-related changes. Five out of twenty two cases (23%) for anemia and 3/11 cases (27%) for thrombocytopenia without otherwise specified pre-bone marrow etiology had uncertain diagnosis after bone marrow biopsy. Conclusion: In about a fifth of patients necessitating a bone marrow, the diagnosis is discordant and can be surprising. It is also worth reporting that in these discordant results, non-hematological causes such as medications, anemia due to chronic diseases or conditions such as cirrhosis or splenomegaly from other etiologies were among the final diagnoses. Interestingly, 20% of the patients with unspecified pre-bone marrow diagnoses such as anemia or thrombocytopenia in our study had an unclear post-bone marrow diagnosis despite undergoing bone marrow biopsy. Our findings are a reminder that the bone marrow exam does not always lead to a definitive diagnosis and the need by exclusion to include in the differential non-hematological etiologies such as nutritional deficiencies, chronic kidney disease or autoimmune disorders. Disclosures No relevant conflicts of interest to declare.

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TIMP-1 and TIMP-2 Levels Are Directly Correlated with Neoangiogenesis and Are Prognostic Factors in Multiple Myeloma Patients.

Blood ◽

10.1182/blood.v114.22.4866.4866 ◽

2009 ◽

Vol 114 (22) ◽

pp. 4866-4866

Author(s):

Luciana Correa Oliveira de Oliveira ◽

Juliana Alves Uzuelli ◽

Ana Paula Alencar de Lima Lange ◽

Barbara Amelia Aparecida Santana-Lemos ◽

Marcia Sueli Baggio ◽

...

Keyword(s):

Multiple Myeloma ◽

Bone Marrow ◽

Overall Survival ◽

Prognostic Factors ◽

Bone Disease ◽

Cox Regression ◽

Conflicts Of Interest ◽

Newly Diagnosed ◽

Significant Difference ◽

The Impact

Abstract Abstract 4866 Background Multiple myeloma (MM) is an incurable malignant disease, characterized by increased angiogenesis in the bone marrow (BM) microenvironment and aberrant BM metabolism. Matrix metalloproteinases (MMP) are a family of zinc-dependent endopeptidases implicated in tumour progression, invasion, metastasis and angiogenesis, via proteolytic degradation of extracellular matrix. MMPs are inhibited by tissue inhibitors of metalloproteinase (TIMP). Although recent studies have implicated MMP 9 in MM bone disease, little is known about the role of the TIMPs. Objectives a) to compare levels of sRANKL, OPG, MMP-2, MMP-9, TIMP-1, TIMP-2, VEGF, bFGF, microvessel density (MVD) between newly diagnosed MM patients and healthy controls; b) to determine the association of these molecules with disease progression, bone disease and neoangiogenesis and c) to evaluate the impact of these variables on survival. Patients and Methods As of July 2009 38 newly diagnosed and untreated multiple myeloma patients were enrolled in the study. The median age was 61years-old (range 39-91) with 24 (63%) males. Patients were diagnosed and categorized according The International Myeloma Working Group criteria and ISS, respectively. Bone involvement was graded according to standard X-ray: patients with no lesions, or with one/ two bones involved or diffuse osteoporosis were classified as low score, whereas patients with lesions in more than two bones or presence of bone fracture were classified as high score. MMP-2 and MMP-9 were determined by PAGE gelatin zymography from plasma as previously described. MMP-9, TIMP-1 and TIMP-2, OPG and sRANKL concentrations were measured by ELISA. The levels of VEGF, bFGF were obtained using cytometric bead array. Ten healthy volunteers were used as controls. Bone marrow MVD measured in hotspots was evaluated in 26 out of 38 patients at diagnosis and 15 patients with Hodgkin Lymphoma stage IA and IIA (used as controls) by staining immunohistochemically for CD34. Comparisons among groups were analyzed by ANOVA and the correlation by the Spearman's correlation coefficient. Cox regression were performed for overall survival (OS) analysis. Results Patients with MM had elevated TIMP-1, TIMP-2 and OPG values compared with controls. No significant difference was found between plasma sRANKL, pro-MMP2, pro-MMP9 and MMP-9 levels. We found that plasma TIMP-1 levels correlated positively with bFGF, VEGF, MVD, beta-2 microglobulin (B2M) and OPG (r: 0.514, p=0,001, r: 0.350, p=0,031; r: 0.610, p<0.0001; r: 0.760, p<0.0001 and r: 0.701, p<0.0001, respectively) and TIMP-2 levels with bFGF, DMV, B2M and OPG (r: 0.512, p=0.002; r: 0.595, p<0.0001; r: 0.587, p<0.0001 and r: 0.552, p<0.0001, respectively). TIMP-1 and TIMP-2 levels correlated with the ISS stage (p<0.0001, p=0.006, respectively). The only variables that correlated with clinical bone disease staging were hemoglobin, B2M and albumin levels, whereas TIMP-1, TIMP-2, bFGF, VEGF and OPG correlated with DMV. On the univariate analyses, age, gender, proMMP2, TIMP-1, TIMP-2, creatinine, B2M and MVD were significantly associated with overall survival. In Cox regression model, TIMP-1, TIMP-2 and B2M levels remained to be significantly associated with OS. In conclusion, our results suggest that TIMP-1 and TIMP-2 levels are strongly associated with neoangiogenesis and are independent prognostic factors in MM. Disclosures No relevant conflicts of interest to declare.

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Association of PML Expression with Chemotherapy-Resistance In Multiple Myeloma Cells

Blood ◽

10.1182/blood.v116.21.2959.2959 ◽

2010 ◽

Vol 116 (21) ◽

pp. 2959-2959

Author(s):

Daisuke Ohgiya ◽

Makoto Onizuka ◽

Hiromichi Matsushita ◽

Naoya Nakamura ◽

Hiroshi Kawada ◽

...

Keyword(s):

Multiple Myeloma ◽

Bone Marrow ◽

Cellular Proliferation ◽

Conflicts Of Interest ◽

Alkylating Agents ◽

Chemotherapy Resistance ◽

Nuclear Body ◽

Rank Test ◽

Significant Difference ◽

The Impact

Abstract Abstract 2959 Background: Although several novel agents have improved the prognosis of patients with multiple myeloma (MM), it still remains an incurable disease because of the difficulty to eradicate MM cells by current therapeutic approaches. Recent studies have revealed that a subset of malignant cells, cancer stem cells, contribute to chemotherapy-resistance in cancer treatment. Promyelocytic leukemia gene product (PML), known as a tumor suppressor through a variety of cellular functions in a nuclear macromolecular structure called the PML nuclear body, has been reported to be responsible for the chemotherapy-resistance by regulating cell cycle in chronic myeloid leukemia. We therefore investigated the impact of PML expression on the cellular proliferation status of MM cells and patients' prognoses. Materials/Methods: Bone marrow clot sections from 48 patients with newly diagnosed MM from Jan 1998 to Dec 2009 before any therapy at diagnosis were obtained, and analyzed, according to appropriate procedure approved by IRB at the Tokai University School of Medicine (Kanagawa, Japan) with written informed consent. They were doubly-stained with a combination of anti-PML/anti-CD138 and anti-Ki67/anti-CD138. For evaluation of the relation between PML status and cellular proliferation, the positive rates of PML and Ki67 in CD138 positive cells were compared. For investigation of the impact of PML expression on the prognosis of MM, the patients were divided into 3 groups, according to the PML positive rates in the CD138 positive cells: negative/low (less than 25 percentile: 12 cases), intermediate (from 25 to 75 percentile: 24 cases) and high (more than 75 percentile: 12 cases). Their overall survivals were compared using log-rank test. Furthermore, the PML positive rates between before and after treatments were compared using paired t-test. Results: The median observation period of 48 cases was 915 days. The median age of the patients was 62.5 (38-76) at diagnosis. All the patients were underwent combination chemotherapies containing alkylating agents as initial therapies. Two and nine patients were underwent allogeneic and autologous stem cell transplantation during the clinical courses, respectively. The numbers of patients of international staging system (ISS) stage I, II and III were 17, 14 and 17 cases. The PML positive rates in each case ranged from 0% to 83.8%. They were not correlated with ISS stages (Spearman r = 0.083) and the Ki67 positive rates (Spearman r = -0.13). The PML positive rates in the negative/low, intermediate and the high groups were less than 22.1%, from 22.1 to 56.6% and more than 56.6%, respectively. No significant difference in overall survival was observed among the 3 groups (p>0.05). However, there were significant differences in two year survival rate when the 3 groups were compared (100%, 85.2% and 54.7%; p=0.015) (Fig. 1). In 13 patients whose bone marrow clot sections were sequentially collected, the PML positive rates after treatments were significantly higher than those at diagnosis (p=0.0042) (Fig. 2). Especially, PML positive rates in all the 3 patients from the negative/low group were progressively increased (0.3 to 82.6%, 14.1 to 100%, 19.0 to 37.5%), and 2 of them died due to disease progression. On the other hand, 2 patients whose PML positive rates decreased after treatment were alive more than 5 years without therapies. Conclusion: Our data indicated that the level of the PML expression at diagnosis was a possible prognostic factor for early course of the disease (2 years after diagnosis). Chemotherapies might induce PML expression in MM cells or select PML positive MM cells. These findings suggest that PML expression presumably reflect chemotherapy-resistance in MM cells. The molecular mechanism of the association is now under investigation. Disclosures: No relevant conflicts of interest to declare.

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Immunophenotypic Response After Allografting In Multiple Myeloma

Blood ◽

10.1182/blood.v122.21.3371.3371 ◽

2013 ◽

Vol 122 (21) ◽

pp. 3371-3371 ◽

Cited By ~ 1

Author(s):

Luisa Giaccone ◽

Lucia Brunello ◽

Roberto Passera ◽

Moreno Festuccia ◽

Milena Gilestro ◽

...

Keyword(s):

Multiple Myeloma ◽

Bone Marrow ◽

Flow Cytometry ◽

Conflicts Of Interest ◽

Univariate Analysis ◽

Conditioning Regimen ◽

First Line ◽

Significant Difference ◽

The Impact

Abstract Background Minimal residual disease (MRD) by multiparameter flow-cytometry recently showed a promising role in predicting outcomes in patients with multiple myeloma. However, data on immunophenotypic response (IR) after allografting are lacking. Aim To evaluate the impact of IR and compare it to conventional complete remission (CR) following allografting in myeloma patients. Methods Sixty-six consecutive patients, median age 54 years (35-66), who underwent an allograft between January 2000 and December 2011 with a follow-up of at least 3 months were included. Disease response was evaluated by serum and urine electrophoresis, and bone marrow aspirate at baseline, 3, 6, 12, 18, 24 months after transplant and yearly thereafter. Skeletal survey or MRI were performed yearly or as clinically indicated (overt relapse or complaints of bone pain). Bone marrow aspirates had to contain at least 13000 cells/µL for flow-cytometry studies and IR was defined as absence of monoclonal plasma-cells detected by 4 or 6-colour staining with the following antibodies: CD38, CD138, CD56, CD19, CD45, cyKappa, cyLambda. CR was defined according to standard criteria (Durie et al, Leukemia 2006; 20:1467-73). Results Conditioning regimen was non-myeloablative 2Gy TBI-based in 55 patients, reduced intensity (fludarabine-melphalan-based) in 10 and myeloablative in 1 patient. Post-grafting immunosuppression consisted of cyclosporine with mycophenolate mofetil or methotrexate. Donors were HLA identical siblings in 58 patients and unrelated in 8. Only 1 patient received bone marrow as source of stem cells. Thirty-five/66 (53%) received the allograft as part of the first line treatment, whereas the remaining 31/66, (47%) were transplanted at relapse. At the time of transplant, 5/66 were both in IR and CR, 16 were only in IR and 4 patients were only in clinical CR. All 21 patients in IR at the time of transplant maintained it, while 26/45 (58%) entered IR after the allograft. Among patients surviving at least 3 months, overall treatment related mortality was 10.6% at 3 years. After a median follow-up of 69 months (range 19-147), the incidence of acute and chronic graft-versus-host disease was 45.6% and 49.3% without significant difference between responsive and non-responsive patients. At follow-up, overall, 24 patients achieved CR and IR (CR/IR group), 21 achieved IR but not CR because of persistence of urine/serum M-component (noCR/IR group), and 21 did not achieve either CR or IR (noCR/noIR group). Interestingly, none achieved CR without IR. Median overall survival (OS) and event-free survival (EFS) in patients who achieved IR were 96 and 55 months versus 36 and 7 months in those who did not (p<0.001). Median OS and EFS were not reached and 59 months in the CR/IR group, 77 and 15 months in the noCR/IR, and 30 and 5 months in the noCR/noIR respectively (p<0.001 for both EFS and OS-fig.1). In univariate analysis, being in the CR/IR group was the only significant predictor for prolonged OS and EFS (p<0.001). Of note, cumulative incidence of extra-medullary disease at first relapse after the allograft was 4% in the CR/IR, 32% in the noCR/IR and 15% in the noCR/noIR groups respectively (p<0.001). Receiving the allograft as first line therapy or later during the disease course did not significantly impact on OS and EFS. Conclusion The achievement of IR confers a favorable impact on OS and EFS after allografting. A higher incidence of extra-medullary in the noCR/IR group (some 30% of our patient cohort) may suggest that myeloma cells escape immune control outside the bone marrow. In this group, imaging studies such as positron emission tomography may clinically be indicated during follow-up to detect early relapse. Disclosures: No relevant conflicts of interest to declare.

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Observations On Eosinophil Ifiltrates in The Bone Marrow Of Patients With Multiple Myeloma. Clinicopathological Correlates

Blood ◽

10.1182/blood.v122.21.5338.5338 ◽

2013 ◽

Vol 122 (21) ◽

pp. 5338-5338

Author(s):

Finella MC Brito-Babapulle ◽

Tanya Cranfield ◽

Robert B Corser ◽

Helen Dignum ◽

Christopher James ◽

...

Keyword(s):

Multiple Myeloma ◽

Bone Marrow ◽

Plasma Cell ◽

Plasma Cells ◽

Conflicts Of Interest ◽

Osteolytic Lesion ◽

Inverse Correlation ◽

Eosinophil Infiltration ◽

Bone Marrow Biopsies ◽

Lytic Lesions

Abstract Mouse eosinophils have been shown in 2011 to be required for the maintenance of long lasting plasma cells in the bone marrow and in maintaining the bone marrow plasma cell microenvironment. Human eosinophils have been shown by Wong et al to support multiple myeloma cell proliferation via a mechanism independent of IL6. We looked at bone marrow biopsies taken from patients who had a paraprotein and in whom a diagnosis of multiple myeloma was suspected. These samples were taken solely for the purposes of diagnosisng multiple myeloma and were retrospectively reviewed from the point of view of degree of eosinophil infiltration and its correlation with tumour load, bone lytic lesions, plasma cell morphology, whether blastic, crystalline inclusions, Mott cells, flame cells and or lymphoplasmacytoid. There were no cases of IGD or E myeloma or osteosclerotic myeloma.Nonsecretory myeloma and cases of light chain myeloma with or without amyloid were included in the series. Biopsies were not performed from osteolytic lesion unless biopsy was necessary to make a diagnosis of myeloma. Myeloma was diagnosed when plasma cell infiltrate was greater than 10% on bone marrow aspirate with a paraprotein and or lytic lesions. Eosinophil infiltration did not correlate with any of the tumour clinicopathological markers but showed an inverse correlation with degree of plasmacytosis. Eosinophils were hardly ever found in marrow aspirates that had over 70% plasma cells. They were usually found in trephine sections of bone marrow in areas where there was Grade I/II fibrosis and were often found in close proximity to focal areas of plasma cell infiltration. Whether eosinophils play a role in preventing or maintaining malignant plasma cell recurrence is currently being studied. Disclosures: No relevant conflicts of interest to declare.

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The Clinical Significance of Argonaute 2 Expression and Angiogenesis in Multiple Myeloma

Blood ◽

10.1182/blood.v126.23.5312.5312 ◽

2015 ◽

Vol 126 (23) ◽

pp. 5312-5312

Author(s):

Qiguo Zhang ◽

Hongyan Wu ◽

Jian Ouyang ◽

Bing Chen ◽

Haibo Dong ◽

...

Keyword(s):

Multiple Myeloma ◽

Bone Marrow ◽

Western Blot ◽

Clinical Significance ◽

Conflicts Of Interest ◽

Negative Group ◽

Bone Marrow Biopsies ◽

Positive Group ◽

Argonaute 2 ◽

Normal Controls

Abstract Objective To assess the argonaute 2 (Ago2) expression in the bone marrow of multiple myeloma (MM) and determine its association with angiogenesis. Methods Fifty-nine MM patients and sixteen normal controls were included. The bone marrow sections were made from plastic (methyl-methacrylate)- embedded bone marrow biopsies. The expression of Ago2 and CD34 in the bone marrow sections was detected by EnVison immunohistochemistry and the microvessel density (MVD) was then assessed. The bone marrow Ago2 expression level was also compared by western blot. The clinical significance of Ago2 expression and MVD in MM was analyzed. Results Western blot analysis indicated that the Ago2 expression was significantly increased in the bone marrow of MM patients as compared with normal controls. As assessed by immunohistochemistry, fifty MM patients were positive for Ago2 and nine patients were negative. However, sixteen normal controls were all negative for Ago2 (χ2=42.586,P<0.001). The β2-microglobulin levels were significantly higher in the Ago2 positive group as compared with the Ago2 negative group (Z=-2.014 , P=0.042). The MVD was significantly higher in bone marrow samples of MM patients compared to the normal controls (7.89±4.88 vs. 2.16±1.32, Z=-3.283 , P<0.001). Analysis of the correlation coefficients showed that Ago2 expression was positively associated with MVD in MM patients (r=0.461, P=0.023). MVD was also significantly higher in the Ago2 positive group than in the Ago2 negative group (Z=-2.449,P=0.014). Conclusion Ago2 expression was found to be significantly increased in the bone marrow of MM patients. Ago2 may take part in the pathogenesis of MM and mediate the angiogenesis. Disclosures No relevant conflicts of interest to declare.

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Bone Marrow-Derived Mesenchymal Stem Cells Inhibit CD8+ t Cell Immune Responses Via PD-1/PD-L1 Pathway in Multiple Myeloma

Blood ◽

10.1182/blood-2020-138418 ◽

2020 ◽

Vol 136 (Supplement 1) ◽

pp. 53-54

Author(s):

Zhaoyun Liu ◽

Fu Mi ◽

Mei Han ◽

Mengyue Tian ◽

Hui Liu ◽

...

Keyword(s):

Stem Cells ◽

Multiple Myeloma ◽

Bone Marrow ◽

T Cells ◽

Mesenchymal Stem Cells ◽

T Cell ◽

Cd8 T Cells ◽

Conflicts Of Interest ◽

Significant Difference ◽

Normal Controls

High expression of the inhibitory receptor programmed death ligand 1 (PD-L1) on tumor cells and tumor stromal cells have been found play a key role in tumor immune evasion in several human malignancies. However, the expression of PD-L1 on bone marrow mesenchymal stem cells (BMSCs) and whether the PD-1/PD-L1 signal pathway is involved in the BMSCs versus T cell immune response in Multiple Myeloma (MM) remain poorly defined. In this study, we explored the expression of PD-L1 on BMSCs from newly diagnosed MM (NDMM) patients and the role of PD-1/PD-L1 pathway in BMSCs-mediated regulation of CD8+T cells. The data showed that the expression of PD-L1 on BMSCs in NDMM patients was significantly increased than that in normal controls (NC) (18.81±1.61% vs. 2.78±0.70 %; P<0.001). Furthermore, the PD-1 expression on CD8+T cells with NDMM patients was significantly higher than that in normal controls (43.22±2.98% vs. 20.71±1.08%; P<0.001). However, there was no significant difference in PD-1 expression of CD4+ T cells and NK cells between NDMM group and NC group. Additionally, the co-culture assays revealed that BMSCs significantly promoted CD8+ T cells apoptosis and suppressed CD8+ T cells function. However, PD-L1 inhibitor effectively reversed BMSCs-mediated suppression in CD8+ T cells. We also found that the combination of PD-L1 inhibitor and pomalidomide can further enhance the killing effect of CD8+ T cells on MM cells. In summary, our findings demonstrated that BMSCs in patients with MM may induce apoptosis of CD8+T cells through the PD-1/PD-L1 axis and inhibit the release of perforin and granzyme B from CD8+ T cells so as to promote the immune escape of MM. Disclosures No relevant conflicts of interest to declare.

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Bone Marrow Fibrosis In Patients With Multiple Myeloma: A New Prognostic Factor For Survival?

Blood ◽

10.1182/blood.v122.21.1946.1946 ◽

2013 ◽

Vol 122 (21) ◽

pp. 1946-1946 ◽

Cited By ~ 2

Author(s):

Tinna Hallgrimsdottir ◽

Anna Porwit ◽

Magnus Björkholm ◽

Eva Rossmann ◽

Hlif Steingrimsdottir ◽

...

Keyword(s):

Multiple Myeloma ◽

Bone Marrow ◽

Plasma Cells ◽

Cox Regression ◽

Conflicts Of Interest ◽

University Hospital ◽

Bone Marrow Fibrosis ◽

Significant Difference ◽

The Difference ◽

Marrow Fibrosis

Abstract Introduction Multiple myeloma (MM) is characterized by the proliferation of plasma cells in the bone marrow and a secretion of monoclonal immunoglobulins. Survival in MM is very variable and multiple factors are known to influence prognosis such as age, ISS stage, and genetic abnormalities. Fibrosis can be found in the bone marrow of MM patients but the literature reporting the incidence of fibrosis and its effect on prognosis is very limited. The purpose of this study was to estimate the incidence of bone marrow fibrosis in MM patients and its effect on survival. Materials and methods Data was collected at the Karolinska University Hospital in Solna, Sweden and information obtained from the hospital's records. We gathered information on all patients diagnosed with MM between 2003 and 2011. All bone marrow reports were reviewed and the presence of bone marrow fibrosis (evaluated using reticulin staining) at diagnosis was recorded. Fibrosis was graded as 1 (mild), 2 (significant) and 3 (advanced), in accordance with WHO 2008 criteria. Patients with fibrosis were paired with patients without fibrosis (matched by sex, birth year, and year of diagnosis). Survival comparing MM patients with and without fibrosis was evaluated using Kaplan-Meier estimate and Cox regression model. Results A total of 586 individuals, 327 males and 259 females, were diagnosed with MM at the Karolinska University Hospital, Solna during 2003 – 2011. Evidence of bone marrow fibrosis was noted in 223 (38%) patients at diagnosis, and 175 had fibrosis grade 1, 33 grade 2, and 15 grade 3. No significant difference was observed between males (N = 135) and females (N = 88) (p = 0.085). Mean age at diagnosis was significantly lower for patients with fibrosis (67.1 years) than in patients without fibrosis (69.7 years) (p = 0.013). Compared with paired patients without fibrosis (N = 217), patients with fibrosis had significantly worse survival (Figure), being 5.0 years vs. 4.4 years, respectively (relative risk (RR)=1.3, 95% confidence interval (CI) 1.00-1.70; p= 0.049). The difference was greatest in male patients and patients younger than 65 years at diagnosis. Survival was worse in patients with advanced fibrosis, 4.5 (95% CI 3.6-6.4) years for grade 1 fibrosis, and 3.0 (95% CI 1.6-NA) years for higher degree of fibrosis. Conclusion In this study, based on almost 600 patients with MM we show that bone marrow fibrosis is common at diagnosis (38%). Importantly, our findings show that the presence of fibrosis was associated with inferior survival. More studies are needed regarding the underlying causes for these findings, including treatment response, treatment-related complications and relation to other known prognostic factors. Disclosures: No relevant conflicts of interest to declare.

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Expression Of Cancer-Testis Antigen In Multiple Myeloma

Blood ◽

10.1182/blood.v122.21.5341.5341 ◽

2013 ◽

Vol 122 (21) ◽

pp. 5341-5341

Author(s):

Jingna Ji ◽

Shangqin Liu ◽

Li He ◽

Chaoping Xu ◽

Guiying Hu ◽

...

Keyword(s):

Multiple Myeloma ◽

Bone Marrow ◽

Healthy Volunteers ◽

Cell Lines ◽

Plasma Cells ◽

Conflicts Of Interest ◽

Clinical Stage ◽

Disease Stage ◽

Significant Difference ◽

Expression Frequency

Abstract In order to detect the CTAs expression in the cell lines and in patients with multiple myeloma (MM). Reverse transcriptase polymerase chain reaction (RT-PCR) detects the mRNA expression of MAGE-C1/CT7, SSX1, SSX2 and SSX4 in MM cell lines RPMI8226 and U266 and patients. Collect clinical MM patients with bone marrow specimens of 25 cases,18 cases of healthy volunteers as a control, The expression frequency in MM patients of CTA gene is parametric statistical analysis with age,gender,the amount of plasma cells,clinical stage and MM type. The CTA members we detected all express in RPMI8226 and U266 cell lines, the expression frequencies of MAGE-C1/CT7 of SSX1, SSX2 and SSX4 in 25 cases bone marrow of MM patients are as follows: 28%（7/25）、80%（20/25）、40%（10/25）、68%（17/25）. 18 cases of healthy volunteers with bone marrow does not express the gene. Four kinds of genes in the bone marrow of patients with simultaneous expression of two or more frequency 80%（20/25）, at least have an expression of the frequency 88%（22/25）. Expression of SSX1 and SSX4 in different disease stage was statistically significant(P <0.05), expression frequency was mainly for patients in the phase Ⅲhigher thanⅠand Ⅱ. Expression of MAGE-C1/CT7 and SSX2 in the period of disease was not statistically significant(P﹥0.05), Detected by the four CTA with age, gender, MM type and the volume of plasma cells was no significant difference(P﹥0.05). It was suggested MAGE-C1/CT7, SSX1, SSX2 and SSX4 gene in MM cell lines RPMI8226 and U266 and MM patients can co-express, while do not express in healthy people. In the expression frequency, SSX1 and SSX4 relate with MM clinical stage,MAGE-C1/CT7 and SSX2 do not relate with clinical stage,the 4 CTAs do not relate with gender,age,clinical type and volume of plasma cells. it provides theoretical support for the CTA vaccine in multiple myeloma immunotherapy. Disclosures: No relevant conflicts of interest to declare.

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