MicroRNA-125b as a valuable predictive marker for outcome after autologous hematopoietic stem cell transplantation.

Background

Many years have passed since autologous hematopoietic stem cell transplantation (AHSCT) or rescue therapy was considered the primary and effective therapeutic approach to multiple myeloma (MM), lymphoma, and other neoplasms [ 1 , 2 ]. Relapse is one of the leading causes of treatment failure after stem cell transplantation [ 3 ]. While some patients get long-term remissions and rarely, even cure, the prognosis for high-risk MM patients remains poor. Treatment response and early relapse after AHSCT can be affected by characteristics associated with patients or diseases, such as weakness, clinical stage, age, cytogenetic abnormalities, and comorbidities [ 4 ]. Early relapse will decrease survival in standard and high-risk MM patients [ 4 ]. The relapsed cases have so heterogeneous pattern of clinical expression. Therefore, a personalized approach should be taken for treatment in transplantation time [ 5 ]. Also, recurrence of Hodgkin lymphoma after AHSCT occurs in 50% of cases in a year [ 6 ]. Relapse after AHSCT in non-Hodgkin lymphoma like follicular lymphoma is common and treatment line, age, graft purging, and myeloablative regimens are considered effective factors in it [ 7 ].

MicroRNAs (miRNAs or miRs) as small non-coding RNAs have critical and wide roles in the expression of genes and also cellular function including cell cycle, growth, differentiation, apoptosis, and so on, either in normal or in the malignant form [ 8 ]. Many of miRNAs have been used as biomarkers for early detection, target therapy, and follow-up in various diseases like lymphoma [ 9 ]. For instance, up-regulation of miR-125b results in down-regulation of its target i.e., SRY-Box Transcription Factor 30 (SOX30) in malignant lymphomas that can be regarded as a useful biomarker, diagnostically and therapeutically [ 10 ]. Poor clinical outcomes are associated with elevated expression of mir-126 in angioimmunoblastic T-cell lymphoma [ 11 ]. miR-150 increases the sensitivity of natural killer/ T (NK/T) cell lymphoma to ionizing radiation through suppression of phosphoinositide 3-kinases/ serine-threonine protein kinas/ mechanistic target of rapamycin kinase (PI3K/AKT/mTOR) pathway [ 12 ] and in MM it has seen that less expression of miR-155 related to resistance to bortezomib [ 13 ]. miRNAs can be packaged in membranous enclosed structures, known as extracellular vesicles (EVs), carried to definite targets via plasma, and protected from environmental factors [ 14 ].

Here, we were going to find a predictive marker for outcomes after AHSCT like relapse or signs of relapse before being evident. In our center, many variables related to patients and microRNA expression are considered in this regard. Given to importance of miRNAs, especially those in EVs, in this study the predictive value of miR-125b, miR-126, miR-150, and miR-155 expression in plasma and EVs, along with many outcome-related data from AHSCT candidates has been evaluated.

Methods

Patients and sample collection

50 MM and lymphoma candidates for AHSCT in the department of bone marrow transplantation center, Taleghani hospital (Tehran, Iran) were selected between September 2021 and August 2022. All protocols and blood sampling were conducted after filling out the informed consent form by patients and confirmation in ethics committee of Hamadan university of medical sciences (Ethic code No: IR.UMSHA.REC.1400.541). Mobilization was done by granulocyte colony-stimulating factor (G-CSF) administration in a steady state. After mobilization and leukocyte count reaches to the appropriate number, for HSCs collection from peripheral blood, Spectra Optia Apheresis System was applied. Lomustine, Etoposide, Cytarabine, Melphalan (CEAM) or Carmustine, Etoposide, Cytarabine, Melphalan (BEAM), and Melphalan/Velcade were conditioning regimens for lymphoma and MM patients, respectively. Patients’ available data was collected from diagnosis to transplantation phase and almost 90 weeks (as the median) follow-up from medical records and new laboratory tests that were done on the patients. 2–4 milliliters of peripheral blood samples were obtained in EDTA containing tube at two times. The first sample was 1–2 days before mobilization and the second sample was 1–2 days after conditioning and before HSCs re-infusion (post conditioning sample).

EVs isolation and confirmation

Based on our previous study [ 15 ], EV isolation was performed by ultracentrifugation. Plasma was isolated by centrifugation at 1000 g for 10 min. 1/1 dilution of plasma with phosphate buffer solution (PBS) was prepared and then centrifugation was performed for 10 min at 2000 g to precipitate dead cells. Subsequently, the supernatant was re-centrifuged for 30 min at 10,000 g to discard cell debris. The obtained supernatant was ultracentrifuged at 100,000 g for 80 min and after the discard of the supernatant, precipitated EVs were diluted by filtered PBS. Then precipitated EVs were filtered through a 0.2 μm filter to remove apoptotic bodies. Finally, ultracentrifugation was done again like before on it. For confirmation of isolated EVs, CD9 and CD63 markers were evaluated by flow cytometry (Dako antibodies, Denmark, and Attune™ NXT Flow Cytometer). Their size was measured by dynamic light scatter (DLS) (Zeta-sizer, Malvern, UK) and the transmission electron microscopy (TEM) technique was done for morphology visualization of them.

Circulatory microRNAs extraction and expression analysis

Expression of miR-125b, miR-126, miR-150, and miR155 in plasma and EVs, before mobilization and post conditioning were analyzed by real time polymerase chain reaction (PCR). The miR extraction was done on MVs/plasma by GeneAll RiboEx™ LS Kit (GeneAll Biotech, south Korea) based on the manufacturer’s instruction. cDNA was synthesized using ExcelRT ™ Reverse Transcriptase kit (SMOBio, South Korea) according to the manufacturer’s instruction and stem-loop primer (Metabion, Germany). In the expression analysis step, real time PCR reaction was performed in the total volume of 15 µl including 7.5 µl of Master mix 2× (Real Q Plus Master Mix, Denmark), 0.5 µl of each primer, 3 µl of the synthesized cDNA, and 3.5 µl of nuclease-free water in Corbett Rotor-Gene 600 thermocycler (Germany). The reactions were done in this condition: initial 95 °C for 15 min, 40 cycles of denaturation at 95 °C for 10s, annealing at 53–54 °C for 15s, extension at 72 °C for the 20s, and 10 min for the melting curve. miR-16 and SNORD47 geometric mean cycle threshold(CT)was used to calculate basal expression or 2 −ΔCT . Sequences of a housekeeping gene, stem-loop, and miRNAs primers were shown in Table 1 .

Table 1 Primers for expression of microRNAs and stem loop for cDNA synthesis miRs Sequences miR-125b (5P) 5-GATGTCCCTGAGACCCTAA-3 miR-126 (3P) 5-GACAGTCGTACCGTGAGTA-3 miR-150 (5P) 5-GTATGTCTCCCAACCCTTG-3 miR-155 (5P) 5-GCTCAGTTAATGCTAATCGTG-3 miR-16 (5P) 5-GACAGTAGCAGCACGTAAAT-3 SNORD47 5-CCAATGATGTAATGATTCTGCC-3 Reverse 5-GAGGAAGAAGACGGAAGAAT-3 Stem loop 5GAAAGAAGGCGAGGAGCAGATCGAGGAAGAAGACGGAAGAATGTGCGTCTCGCCTTCTTTCHVMHNN-3

Patients follow up

After 16–130 weeks, almost 90 weeks (as the median), all of the patients were followed about their outcomes. Factors including complete blood count (CBC) parameters, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), urea, creatinine, lactate dehydrogenase (LDH), calcium, alkaline phosphatase, total protein, gamma globin, M protein spike, β2 macroglobulin, lytic lesion in bone, plasma cell percent in bone marrow, and clinical features like lymphadenopathy, bone pain, weakness, headache, and fever as outcomes were evaluated and recorded from medical reports and new laboratory tests that were done on the patients. These factors are related to remission and relapse.

Statistical analysis

All of the collected data from diagnosis to transplantation and follow-up time were analyzed in Poisson multivariate analysis to find predictive markers for all outcome factors using South Texas Art Therapy Association STATA software (College Station, Texas, USA, V14). Significant hazard ratio (HR) and p value show how often relapse happen over time in a dysregulated parameter. Evaluation of the predictive value for variables in the prediction of outcomes was analyzed by receiver-operating characteristic (ROC) analysis. In addition, Kaplan-Meier was used to estimate the cumulative incidence (CI) of outcomes and relapse free survival (RFS) analysis by effective variables. p value less than 0.05 was considered statistically significant.

Results

Patients’ data

50 candidates of AHSCT, with a mean and standard deviation of age 42.4 ± 14.6 years and 50% of them were female, were incorporated into the study. The median time for the engraftment of neutrophils and platelet was 11 days after AHSCT. Demographic data, CD34 + cell count/kg harvested by mobilization and apheresis, mobilization day number, the total dose of G-CSF, neutrophil and platelet count at apheresis day, and other AHSCT-related data were recorded (Table 2 ). The median follow-up was 90 weeks (range 16–130 weeks) and nine patients relapsed. In Table 2 , other recorded variables as outcomes were shown.

Table 2 Participants criteria (N = 50) Variables at the AHSCT time Outcome variables Features Median (Rang) / Frequency (%) Features Median (Rang) / Frequency (%) Age (year) 40.5 (18–70) Hemoglobin (g/dl) 12.3 (8.8–16.2) Gender WBC (×10 3 /µl) 5.03 (2.8–10.2) Male 25 (50%) PLT (×10 3 /µl) 178 (133–295) Female 25 (50%) ESR (mm/hours) 13.5 (2–82) Diagnosis LDH (U/L) 287 (148–450) Multiple myeloma 24 (48%) Ca (mg/dl) 9.3 (7.8–13) Hodgkin lymphoma 15 (30%) Cr (mg/dl) 1.03 (0.6-2.0) Non-Hodgkin lymphoma 11(22%) ALP (IU/L) 102 (51–373) BMI after transplantation 27.1 (14.8–42.1) Urea (mg/dl) 18 (11–23) Relapse 9 (18%) Mobilization day number 6 (5–9) Positive lytic lesion (In MM patients) 2 (8%) Total dose of infused G-CSF for mobilization (µg) 3750 (1500–7200) Positive M protein spike 4 (16%) WBC count at start day of mobilization (×10 3 /µl) 6.05 (2.4–22.1) Positive radiologic signs in lymphoma patients 6 (23%) Last injection of G-CSF to apheresis (hour) 6 (1–22) Neutrophil count on apheresis day (×10 3 /µl) 35.13 (10.7–94.5) Apheresis duration (min) 377.5 (270–520) Total processed blood in apheresis (Littr) 17.97 (10.4–25.5) WBC count in apheresis product (×10 8 /kg) 10.9 (4.2–26.2) MNC count in apheresis product (×10 8 /kg) 6.20 (3.3–17.5) CD34 + cell count/kg (×10 6 /kg) 3.5 (1-15.4) CD34 + in PAPBS (/µl) 35 (4.1–116) Platelet engraftment day 11 (9–19) Platelet count at engraftment day (×10 3 /µl) 28 (20–110) BMI : Body mass index, WBC : White blood cell, MNC : Mono nuclear cell, PAPBS : Pre-apheresis peripheral blood sample, ESR : Erythrocyte sedimentation rate, LDH : Lactate dehydrogenase, Ca : Calcium, Cr : Creatinine, ALP : Alkaline phosphatase

Isolated EV’s characterization

Expression of CD9 and CD63 on EVs were verified by flow cytometry (Fig. 1 A). DLS analysis of isolated EVs indicated that the majority of EVs were ~ 100 nm (Fig. 1 B).TEM images confirmed the morphological criteria of EVs (Fig. 1 C).

Fig. 1 Isolated EVs confirmation. A : Flow cytometry for CD9 and CD63 with 87.4% and 52.1% positivity, respectively. B : DLS results indicated majority of EVs were ~ 100 nm. C : TEM images were in accordance with DLS results in term of size. EVs : Extracellular vesicles, DLS : Dynamic light scatter, TEM : Transmission electron microscopy

The microRNAs expression

Basal expression of four circulatory miRNAs including miR-125b, miR-126, miR-150, and miR155 in plasma and EVs, before mobilization and post conditioning were analyzed. Significantly higher expression of miR-125b was detected in EVs before mobilization in comparison with plasma (p value < 0.001) (Fig. 2 A). Both miR-150 and miR-155 in EVs were significantly higher than plasma post conditioning (p value < 0.05) (Fig. 2 B). There was no significant difference between before mobilization and post conditioning miRNAs, except miR-150 which was higher before than post conditioning (p value < 0.01) (Fig. 2 B).

Fig. 2 Expression of miRNAs in EVs and plasma. Expression of miR-125b, miR-126, miR-150, and miR155 in plasma and EVs, before mobilization and post conditioning were analyzed by real time polymerase chain reaction. *, ** and *** p value < 0.05, < 0.01 and < 0.001. EVs : Extracellular vesicles, BE : Before mobilization EVs, BP : Before mobilization plasma, PE : Post conditioning EVs, PP : Post conditioning plasma

Multi-variate and ROC analysis results

COX and Poisson regression (multi-variate) analysis were done on all variables and all outcomes that were shown in Table 3 . These covariates which could be related to outcomes like relapse, LDH, and ESR, were obtained from available medical records and tests and we wanted to evaluate these. There were valuable predictive markers for relapse, high LDH (> 280 U/L), and high ESR (> 25 mm/hour) as outcome variables (Table 3 ). miR-125b expression level before mobilization associated with relapse, high LDH, and high ESR level. Based on the results, miR-125b before mobilization was associated with all three outcome parameters. miR-125b before mobilization in EVs can predict relapse with HR: 2.23 and p value: 0.01, and in plasma with HR: 6.7 and p value < 0.001 (Table 3 ). As shown in Table 3 , some other variables were significantly associated with relapse, high LDH, and high ESR through multivariate analysis. We have analyzed the predictive value of miR-125b in the prediction of the three outcomes by ROC that was shown in Fig. 3 A-E. miR-125b level before mobilization in both EVs and plasma was a convenient predictor for relapse, high ESR, and high LDH after AHSCT (Fig. 3 A-E) (p value: 0.0001, 0.024, 0.001, 0.018, and 0.002).

Fig. 3 Receiver-operating characteristic (ROC) analysis. ROC curves indicated powerful predictive value of miR-125b in EVs and plasma for relapse (A), in EVs for high ESR (B) in plasma for high ESR (C), in EVs for high LDH (D), and in plasma for high LDH (E). EVs : Extracellular vesicles, ESR : Erythrocyte sedimentation rate, LDH : Lactate dehydrogenase

Table 3 Predictive variable for relapse, LDH, and ESR as AHSCT outcomes by multivariate analysis. COX regression for relapse and Poisson regression for LDH and ESR Relapse LDH ESR Variables HR (95% CI) P value Variables Beta P value Variables Beta P value miR-125b BE 2.23 (2.12–2.78) 0.01 Intercept 6.65 < 0.0001 Intercept 2.01 0.044 miR-125b BP 6.7 (6.41–7.02) < 0.001 miR-125b BE 4.79 < 0.0001 miR-125b BE 5.65 < 0.0001 Age 0.24 (0.20–1.02) 0.426 miR-125b BP 8.52 < 0.0001 miR-125b BP 2.41 0.015 Engraftment day 1.04 (0.74–1.91) 0.091 Age 0.35 0.72 Age 1.11 0.267 Diagnosis to transplantation 0.72 (0.57–1.22) 0.372 Weight 2.45 0.064 Weight 1.65 0.098 G-CSF dose for mobilization 0.8 (0.07–1.8) 0.263 Diagnosis to transplantation 1.83 0.067 Chemotherapy Courses 1.48 0.139 CD34 + cell count/kg 1.01 (0.04–1.43) 0.117 Mobilization Days 4.46 < 0.1 WBC count in apheresis 0.92 0.356 G-CSF dose for mobilization 6.28 0.1 MNC count in apheresis 1.74 0.081 Pre-apheresis CD34 + cell count 1.78 0.074 CD34 + cell count/kg 2.42 0.15 Total processed blood in Apheresis 3.64 0.12 Conditioning days 1.35 0.177 MNC count in apheresis 4.01 0.08 Engraftment day WBC count 1.97 0.068 CD34 + cell count/kg 3.02 0.0026 Engraftment day PLT count 1.63 0.103 Conditioning days 2.07 0.38 Engraftment Day 2.34 0.19 Engraftment day 4.50 < 0.0001 miR-155 PE 0.69 0.489 miR-155 BP 1.26 0.207 miR-126 BP 1.17 0.07 miR-126 PP 0.09 0.926 ESR : Erythrocyte sedimentation rate, LDH : Lactate dehydrogenase, EVs : Extracellular vesicles, HR : Hazard ratio, BE : Before mobilization EVs, BP : Before mobilization plasma, PE : Post conditioning EVs, PP : Post conditioning plasma, WBC : White blood cell, MNC : Mono nuclear cell, G-CSF : Granulocyte colony stimulating factor

RFS and CI of outcomes

RFS and CI of high LDH (> 280 U/L), and high ESR (> 25 mm/hour) based on miR-125b expression before mobilization was evaluated by the Kaplan-Meier test (Fig. 4 A-F). RFS analysis show increased incidence of relapse (decrease in RFS) after AHSCT increases in miR-125b expression ≥ 2.5 in plasma and EVs, significantly (p value: 0.006 and 0.0003) (Fig. 4 A and B). Based on CI, with an increase of miR-125b expression, high LDH, and high ESR will be increased. More LDH after AHSCT was significantly associated with miR-125b expression of more than 2.1 in plasma (p value: 0.04 and 0.03) (Fig. 4 C and D). Also, increasing miR-125b expression by more than 2.5 in plasma and EVs, resulted in a significantly increasing in ESR after AHSCT (p value: 0.04 and 0.0003) (Fig. 4 E and F).

Fig. 4 Relapse free survival (RFS) analysis and cumulative incidence by Kaplan-Meier test. Decreasing RFS is associated with increased expression of miR-125b ( A & B ), and high LDH ( C & D ) and high ESR ( E & F ) associated with increased expression of miR-125b (more than cut off). BE : Before mobilization EVs, BP : Before mobilization Plasma. p values A : 0.006, B : 0.0003, C : 0.04, D : 0.03, E : 0.04 and F : 0.0003

Discussion

Therapeutic options for relapsed Hodgkin and non-Hodgkin lymphoma are so limited, except allogeneic HSCT with some challenges like graft versus host disease [ 16 ]. There is no standard treatment for relapsed MM after AHSCT [ 17 ]. So, the detection of predicting factors for relapse can be applicable to the prevention of it and therapy modification at AHSCT time [ 18 ]. Also, there are some strategies after AHSCT for improving the outcomes, if physicians are notified in time by some markers [ 18 ]. In our study, circulatory miRNAs and many variables were evaluated to their probable predictive potential of AHSCT and relapse-related outcomes. We found that circulatory miR-125b expression level in EVs and plasma is a strong predictor for relapse after AHSCT and also, LDH and ESR are as informative factors of risk, prognosis, and relapse of MM and lymphoma. In MM patients with upregulated miR-125b, event-free survival is shorter significantly [ 19 ]. miR-125b reduces the metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) gene expression via neurogenic locus notch homolog protein 1 (NOTCH1) signaling pathway in MM cells, results in more progressiveness of MM and it can be a therapeutic target [ 20 ]. Indeed miR-125b, as an onco-miR, targets tumor suppressors including long non-coding RNA named insulin-like growth factor 1 receptor antisense imprinted non-protein coding RNA (IRAIN) and PH domain and leucine-rich repeat protein phosphatase 2 (PHLPP2) that finally elevates AKT signaling pathway and myeloma cell progression [ 20 , 21 ]. However, miR-125b is a multifunctional miRNA [ 22 ] with a positive effect on dexamethasone [ 23 ] and stress adaptation induced myeloma cell death [ 24 ]. Due to the high expression of miR-125b in EVs [ 25 ], its up-regulation in serum associated with Rituximab, Cyclophosphamide, Hydroxy daunorubicin, Vincristine, and Prednisone (R-CHOP) resistance, further relapse, and diminished survival in diffuse large B cell lymphoma patients [ 26 ]. Nuclear factor kappa-light-chain-enhancer of activated B cells (NFkB) activity in B cell lymphoma increases by miR-125b that targets the tumor necrosis factor alpha-induced protein 3, thereby facilitating the cancer progression and cell proliferation [ 27 , 28 ]. Totally, silencing of miR-125b is required for normal hematopoietic stem cell development toward B lymphocyte and neoplasia suppression [ 29 ]. These published studies indicate that mir-125b induces the progression and relapse of MM and lymphoma. Based on our hypothesis, circulatory miR-125b expression level might be able to the prediction of relapse after AHSCT. We introduce miR-125b as a potent predictor of relapse after AHSCT, and also a good predictor of LDH and ESR. Although high LDH is known as a bad prognosis marker in newly diagnosed MM patients [ 30 ], it is a valuable informative parameter about outcomes and survival after allogeneic HSCT for MM [ 31 ]. LDH concentration should be considered as an applicable variable for risk stratification of MM [ 32 ], as a high probability of relapse after two years of AHSCT in patients with a high level of LDH is reported [ 33 ]. In addition, high concentrations of calcium and LDH (> 240 U/L) are criteria for stage III of the international staging system with elevated relapse risk of MM [ 33 ]. In the study, we considered high LDH as more than 280 U/L. Relapse and mortality after AHSCT is predicted by increased LDH (over a period of three months) in patients with lymphoma [ 34 , 35 ].Through LDH along with ESR, survival analysis and monitoring of therapy in MM and Hodgkin lymphoma will be reliable, respectively [ 36 , 37 ]. Elevated ESR after treatment like radiotherapy, chemotherapy, and HSCT is associated with relapse of Hodgkin lymphoma [ 38 , 39 ]. So, evaluation of LDH and ESR can be valuable for the prediction of relapse of MM and lymphoma after AHSCT. Not only the evaluation of these parameters by themselves, weeks after AHSCT but also the measurement of associated parameters like miRNAs at the time of AHSCT could be informative about relapse and its predictive markers (LDH and ESR). Indeed miR-125b is able to prediction of relapse much earlier than LDH and ESR [ 40 ]. According to the effect of miR-125b on MM and lymphoma, administration of miR-125b antagomir inhibits cell proliferation, tumor growth, and neoplasm relapse [ 40 ].

Conclusion

As a conclusion circulatory miR-125b is associated with the risk of relapse and relapse-related parameters in MM and lymphoma patients. Monitoring of the circulatory level of miR-125b at the AHSCT time predicts the treatment response and helps to do additional strategies for the prevention of relapse.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Acknowledgements

This work was founded by Hamadan University of Medical Sciences. Thank Iranian biophysics and biochemistry center for TEM technical support.

Authors’ contributions

M.R. and F.A. wrote the first main manuscript text. MH.M. prepared figures. A.H. collected and analyzed the data. All authors reviewed the manuscript.

Funding

This study was founded by Hamadan University of Medical Sciences, Ethic Code: Ethic code No: IR.UMSHA.REC.1400.541, grant No: 140008186756.

Availability of data and materials

The data have included in the text and documented/reserved with the corresponding author.

Code availability

Not applicable.

Declarations

Competing interests

All authors state that there is no conflict of interest.

Ethics approval

All procedures were in accordance with the ethical standards of the institutional and/or national research committee. This study was confirmed in ethic committee of Hamadan university of medical sciences (Ethic code No: IR.UMSHA.REC.1400.541).

Consent to participate

All protocols and blood sampling were conducted after filling out the informed consent form by patients.

Consent for publication

Not applicable.

Abbreviations

Autologous hematopoietic stem cell transplantation

Multiple myeloma

MicroRNAs

SRY-box transcription factor 30

Natural killer/ T cell

Phosphoinositide 3-kinases/ Serine-threonine protein kinas/ Mechanistic target of rapamycin kinase

Granulocyte colony stimulating factor

Lomustine, Etoposide, Cytarabine, Melphalan

Carmustine, Etoposide, Cytarabine, Melphalan

Phosphate buffer solution

Dynamic light scatter

Transmission electron microscopy

Polymerase chain reaction

Cycle threshold

Complete blood count

Erythrocyte sedimentation rate

C-reactive protein

Lactate dehydrogenase

South texas art therapy association

Hazard ratio

Receiver-operating characteristic

Cumulative incidence

Body mass index

White blood cell

Mono nuclear cell

Count per kilogram

Pre-apheresis peripheral blood sample

Calcium

Creatinine

Alkaline phosphatase

Before mobilization EVs

Before mobilization plasma

Post conditioning EVs

Post conditioning plasma

Metastasis associated lung adenocarcinoma transcript 1

Neurogenic locus notch homolog protein 1

Rituximab, Cyclophosphamide, Hydroxy daunorubicin, Vincristine, and Prednisone

Nuclear factor kappa-light-chain-enhancer of activated B cells

Insulin like growth factor 1 receptor antisense imprinted non-protein coding RNA

PH domain and leucine rich repeat protein phosphatase 2

Serine-threonine protein kinas

References

1. Vaxman I Visram A Kumar S Dispenzieri A Buadi F Dingli D Autologous stem cell transplantation for multiple myeloma patients aged ≥ 75 treated with novel agents Bone Marrow Transplant 2021 56 5 1144 50 10.1038/s41409-020-01159-9 33273658

2. Wudhikarn K Johnson BK Villasboas JC Paludo J Inwards DJ Porrata LF Improvement in outcomes of autologous stem cell transplant in patients with Lymphoma Older Than 70 years: the significance of age in 2020s? Blood 2021 138 2908 10.1182/blood-2021-144507

3. Chen Y-B McCarthy PL Hahn T Holstein SA Ueda M Kröger N Methods to prevent and treat relapse after hematopoietic stem cell transplantation with tyrosine kinase inhibitors, immunomodulating drugs, deacetylase inhibitors, and hypomethylating agents Bone Marrow Transplant 2019 54 4 497 507 10.1038/s41409-018-0269-3 30038353

4. Corre J Montes L Martin E Perrot A Caillot D Leleu X Early relapse after autologous transplant for myeloma is associated with poor survival regardless of cytogenetic risk Haematologica 2020 105 9 e480 10.3324/haematol.2019.236588 33054068 PMC7556617

5. Alegre A Granda A Martínez-Chamorro C Díaz-Mediavilla J Martínez R García-Laraña J Different patterns of relapse after autologous peripheral blood stem cell transplantation in multiple myeloma: clinical results of 280 cases from the Spanish Registry Haematologica 2002 87 6 609 14 12031917

6. Crump M Management of Hodgkin lymphoma in relapse after autologous stem cell transplant ASH Educ Program Book 2008 2008 1 326 33 10.1182/asheducation-2008.1.326 19074105

7. Kornacker M Stumm J Pott C Dietrich S Süssmilch S Hensel M Characteristics of relapse after autologous stem-cell transplantation for follicular lymphoma: a long-term follow-up Ann Oncol 2009 20 4 722 8 10.1093/annonc/mdn691 19179546

8. Asgarpour K Shojaei Z Amiri F Ai J Mahjoubin-Tehran M Ghasemi F Exosomal microRNAs derived from mesenchymal stem cells: cell-to-cell messages Cell Commun Signal 2020 18 1 1 16 10.1186/s12964-020-00650-6 32917227 PMC7488404

9. Han B Wang S Zhao H MicroRNA-21 and microRNA-155 promote the progression of Burkitt’s lymphoma by the PI3K/AKT signaling pathway IJ Clin Exp Pathol 2020 13 1 89 PMC7013371 32055277

10. Zhan C Wang T You H Si C Different expressions of miR-125b and SOX30 in malignant lymphomas and their significance J BUON 2018 23 4 1179 84 30358229

11. Lone W Bouska A Sharma S Amador C Saumyaranjan M Herek TA Genome-wide miRNA expression profiling of molecular subgroups of peripheral T-cell lymphoma Clin Cancer Res 2021 27 21 6039 53 10.1158/1078-0432.CCR-21-0573 34426436 PMC8563427

12. Wu SJ Chen J Wu B Wang YJ Guo KY MicroRNA-150 enhances radiosensitivity by inhibiting the AKT pathway in NK/T cell lymphoma J Exp Clin Cancer Res 2018 37 1 1 10 10.1186/s13046-017-0639-5 29386059 PMC5793389

13. Amodio N Gallo Cantafio ME Botta C Agosti V Federico C Caracciolo D Replacement of miR-155 elicits tumor suppressive activity and antagonizes bortezomib resistance in multiple myeloma Cancer 2019 11 2 236 10.3390/cancers11020236 PMC6406286 30781685

14. Ma Y Li C Huang Y Wang Y Xia X Zheng CJ Exosomes released from neural progenitor cells and induced neural progenitor cells regulate neurogenesis through miR-21a Cell Commun Signal 2019 17 96 10.1186/s12964-019-0418-3 31419975 PMC6698014

15. Rafiee M Allahbakhshian FM Amiri V Hajifathali A Gharehbaghian A Mohammadi MH Circulatory miR–155 correlation with platelet and neutrophil recovery after autologous hematopoietic stem cell transplantation a multivariate analysis Int J Hematol 2021 114 2 235 45 10.1007/s12185-021-03154-2 33895969

16. Smith SM van Besien K Carreras J Bashey A Cairo MS Freytes CO Second autologous stem cell transplantation for relapsed lymphoma after a prior autologous transplant Biol blood Marrow Transplant 2008 14 8 904 12 10.1016/j.bbmt.2008.05.021 18640574 PMC3353768

17. Malard F Harousseau J Mohty M Multiple myeloma treatment at relapse after autologous stem cell transplantation: a practical analysis Cancer Treat Rev 2017 52 41 7 10.1016/j.ctrv.2016.11.005 27888768

18. Sureda A André M Borchmann P da Silva MG Gisselbrecht C Vassilakopoulos TP Improving outcomes after autologous transplantation in relapsed/refractory Hodgkin lymphoma: a european expert perspective BMC Cancer 2020 20 1 1 14 10.1186/s12885-020-07561-2 PMC7657361 33172440

19. Jiang Y Luan Y Chang H Chen G The diagnostic and prognostic value of plasma microRNA–125b–5p in patients with multiple myeloma OncolLett 2018 16 3 4001 7 10.3892/ol.2018.9128 PMC6096151 30128020

20. Gao D Xiao Z Li HP Han DH Zhang YP The mechanism study of miR-125b in occurrence and progression of multiple myeloma Cancer Med 2018 7 1 134 45 10.1002/cam4.1181 29214735 PMC5773953

21. Jiang Y Ding J Li J Chen G Effects of microRNA–125b on multiple myeloma cell growth in vitro and in vivo Oncol Rep 2018 40 5 2864 75 30226579 10.3892/or.2018.6668

22. Wang Y Wei Y Fan X Zhang P Wang P Cheng S MicroRNA-125b as a tumor suppressor by targeting MMP11 in breast cancer ThoracCancer 2020 11 6 1613 20 10.1111/1759-7714.13441 PMC7262928 32291953

23. Murray M Rushworth SA Zaitseva L Bowles KM MacEwan DJ Attenuation of dexamethasone-induced cell death in multiple myeloma is mediated by miR-125b expression Cell Cycle 2013 12 13 2144 53 10.4161/cc.25251 23759586 PMC3737316

24. Misso G Zarone MR Lombardi A Grimaldi A Cossu AM Ferri C miR-125b upregulates miR-34a and sequentially activates stress adaption and cell death mechanisms in multiple myeloma Mol Ther Nucleic Acids 2019 16 391 406 10.1016/j.omtn.2019.02.023 31009917 PMC6479071

25. Feng Y Zhong M Zeng S Wang L Liu P Xiao X Exosome-derived miRNAs as predictive biomarkers for diffuse large B-cell lymphoma chemotherapy resistance Epigenomics 2019 11 1 35 51 10.2217/epi-2018-0123 30211623

26. Yuan WX Gui YX Na WN Chao J Yang X Circulating microRNA-125b and microRNA-130a expression profiles predict chemoresistance to R-CHOP in diffuse large B-cell lymphoma patients Oncol Lett 2016 11 1 423 32 10.3892/ol.2015.3866 26870228 PMC4726930

27. Kim S-W Ramasamy K Bouamar H Lin A-P Jiang D Aguiar RC MicroRNAs miR-125a and miR-125b constitutively activate the NF-κB pathway by targeting the tumor necrosis factor alpha-induced protein 3 (TNFAIP3, A20) PNAS 2012 109 20 7865 70 10.1073/pnas.1200081109 22550173 PMC3356650

28. Peng B Theng PY Le MT Essential functions of miR-125b in cancer Cell Prolif 2021 54 2 e12913 10.1111/cpr.12913 33332677 PMC7848968

29. Li G So AY-L Sookram R Wong S Wang JK Ouyang Y Epigenetic silencing of miR-125b is required for normal B-cell development Blood 2018 131 17 1920 30 10.1182/blood-2018-01-824540 29555645 PMC5921965

30. Gu Y Yuan Y-H Xu J Shi Q-L Qu X-Y Guo R High serum lactate dehydrogenase predicts an unfavorable outcome in chinese elderly patients with multiple myeloma Oncotarget 2017 8 29 48350 10.18632/oncotarget.16237 28418851 PMC5564653

31. Shouval R Teper O Fein JA Danylesko I Shem Tov N Yerushalmi R LDH and renal function are prognostic factors for long-term outcomes of multiple myeloma patients undergoing allogeneic hematopoietic stem cell transplantation Bone Marrow Transplant 2020 55 9 1736 43 10.1038/s41409-020-0829-1 32066863

32. Dimopoulos MA, Kastritis E, Roussou M, Gkotzamanidou M, Migkou M, Gavriatopoulou M, et al. Elevated serum lactate dehydrogenase (LDH) should be included among the variables which define high risk multiple myeloma. Am Society Hematol; 2010.

33. Huang B, Li J, Lu J, Xiao Y, Zhao Y, Huang H. Scoring system for predicting risk of relapse in patients with multiple myeloma within two years after stem cell transplantation. Am Society Hematol Washington, DC; 2017.

34. Sorror M Storer B Gopal A Holmberg L Sandmaier B Bensinger W Comorbidity, Lactate dehydrogenase (LDH), and Chemosensitivity are independent predictors of mortality after autologous hematopoietic cell transplantation (HCT) for patients (pts) with lymphoma Blood 2007 110 11 616 10.1182/blood.V110.11.616.616

35. William BM Bongu NR Bast M Bociek RG Bierman PJ Vose JM The utility of lactate dehydrogenase in the follow up of patients with diffuse large B-cell lymphoma Rev bras hematol hemoter 2013 35 189 91 23904809 10.5581/1516-8484.20130055 PMC3728132

36. Alexandrakis M Passam F Ganotakis E Sfiridaki K Xilouri I Perisinakis K The clinical and prognostic significance of erythrocyte sedimentation rate (ESR), serum interleukin-6 (IL‐6) and acute phase protein levels in multiple myeloma Clin Lab Haematol 2003 25 1 41 6 10.1046/j.1365-2257.2003.00492.x 12542441

37. Bien E Balcerska A Serum Soluble Interleukin-2 receptor, Beta2‐Microglobulin, Lactate dehydrogenase and erythrocyte sedimentation rate in children with Hodgkin’s lymphoma Scand J immunol 2009 70 5 490 500 10.1111/j.1365-3083.2009.02313.x 19874554

38. Friedman S Henry-Amar M Cosset JM Carde P Hayat M Dupouy N Therapeutic implications and sites of relapse predicted by elevated posttherapy erythrocyte sedimentation rate in early stage Hodgkin disease Am J hematol 1991 37 4 253 7 10.1002/ajh.2830370408 1858782

39. Henry-Amar M Friedman S Hayat M Somers R Meerwaldt JH Carde P Erythrocyte sedimentation rate predicts early relapse and survival in early-stage Hodgkin disease Ann Inter Med 1991 114 5 361 5 10.7326/0003-4819-114-5-361 1992877

40. Zheng Z Qu J-Q Yi H-M Ye X Huang W Xiao T MiR-125b regulates proliferation and apoptosis of nasopharyngeal carcinoma by targeting A20/NF-κB signaling pathway Cell Death Disease 2017 8 6 e2855 e 10.1038/cddis.2017.211 28569771 PMC5520883