Prognostic Value of Serum Paraprotein Response Kinetics in Patients With Newly Diagnosed Multiple Myeloma

Response kinetics is not well-established as a prognostic marker in multiple myeloma (MM). We developed a mathematical model to assess the prognostic value of serum monoclonal component (MC) response kinetics during 6 induction cycles in 373 newly diagnosed MM patients. The model calculated a resistance parameter that reflects the stagnation in the response after an initial descent, dividing the patients into two kinetics categories with significantly different progression-free survival (PFS). Introduction: Response kinetics is a well-established prognostic marker in acute lymphoblastic leukemia. The situation is not clear in multiple myeloma (MM) despite having a biomarker for response monitoring (monoclonal component [MC]). Materials and Methods: We developed a mathematical model to assess the prognostic value of serum MC response kinetics during 6 induction cycles, in 373 NDMM transplanted patients treated in the GEM2012Menos65 clinical trial. The model calculated a resistance parameter that reflects the stagnation in the response after an initial descent. Results: Two patient subgroups were defined based on low and high resistance, that respectively captured sensitive and refractory kinetics, with progression-free survival (PFS) at 5 years of 72% and 59% (HR 0.64, 95% CI 0.44-0.93; P =.02). Resistance significantly correlated with depth of response measured after consolidation (80.9% CR and 68.4% minimal residual disease negativity in patients with sensitive vs. 31% and 20% in those with refractory kinetics). Furthermore, it modulated the impact of reaching CR after consolidation; thus, within CR patients those with refractory kinetics had significantly shorter PFS than those with sensitive kinetics (median 54 months vs. NR; P =.02). Minimal residual disease negativity abrogated this effect. Our study also questions the benefit of rapid responders compared to late responders (5-year PFS 59.7% vs. 76.5%, respectively [P <.002]). Of note, 85% of patients considered as late responders were classified as having sensitive kinetics. Conclusion: This semi-mechanistic modeling of M-component kinetics could be of great value to identify patients at risk of early treatment failure, who may benefit from early rescue intervention strategies. (C) 2022 The Authors. Published by Elsevier Inc

Trasplante de células mesenquimales fucosiladas en el tratamiento de la osteoporosis. Estudio preclínico y prueba de concepto

Objetivos: La osteoporosis es una enfermedad ósea definida por la pérdida de masa ósea generalizada con un elevado riesgo de fracturas óseas asociado a una alta morbimortalidad. Las Células Stem Mesenquimales (CSM) de médula ósea (MO) pueden representar un tratamiento óptimo para la osteoporosis debido a su capacidad innata de diferenciación a osteoblastos. Aunque el injerto de las CSM ha sido demostrado, su tropismo óseo es bajo cuando se administran de forma intravenosa, imprescindible para el tratamiento de esta enfermedad sistémica. Su tropismo depende de la interacción de sus receptores de superficie con los que se encuentran en las células endoteliales. La proteína E-selectina es inducida en el endotelio de la microvasculatura de tejidos inflamatorios en respuesta a citoquinas y es constitutivamente expresada en las células endoteliales de MO. La glicoforma de CD44 (HCELL), un potente ligando de E y L-selectina, es expresada de forma innata por las Células Stem Hematopoyéticas (CSH). HCELL es la molécula clave para el reclutamiento celular hacia la MO. Las CSM expresan CD44 pero no HCELL ni otros ligandos de E-selectina de forma constitutiva. La fucosilación (adición de una fucosa) ex vivo en posición α1,3 del antígeno CD44 mediante la enzima fucosiltransferasa VI (FTVI) produce HCELL en las CSM, incrementando su afinidad por E-selectina y por tanto su osteotropismo. EL objetivo de este estudio preclínico es evaluar la seguridad y la eficacia de las CSM humanas fucosiladas en un modelo murino inmunocomprometido (NOD/SCID). Métodos: 31 ratones NOD/SCID fueron randomizados a recibir de forma intravenosa: 1x106 CSM fucosiladas (n=13), 1x106 CSM (n=14), o salino (n=4). Las toxicidades fueron evaluadas por un score clínico, peso y un examen histológico (corazón, pulmón, hígado, bazo, riñón, gónadas, cerebro, hueso y MO). Se realizó estudio de biodistribución mediante detección de los genes β-actina y β2-microglobulina humanas. La integridad genética fue evaluada por cariotipo. Se realizó inmunohistoquímica con un anticuerpo policlonal anti-Osteocalcina humana para valorar eficacia. Las células osteocalcina positivas fueron identificadas por un precipitado citoplasmático de color marrón oscuro. Resultados: No se produjo ningún fallecimiento inesperado y ningún ratón mostró toxicidad aguda. Un ratón en el brazo de salino presentó toxicidad crónica leve (piloerección y cambios moderados en el comportamiento). La histología de corazón, hígado, bazo, riñón, cerebro, hueso y MO fue normal en todos los ratones. Se objetivaron áreas localizadas inflamatorias en los pulmones de un 15%, 42% y 25% de los ratones infundidos con CSM fucosiladas, CSM y salino respectivamente sin diferencias significativas (p=0.28). La biodistribución fue normal en todos los ratones con la excepción de un ratón (infundido con CSM sin fucosilar) que mostró expresión de ARN humano (β-actina and β2-microglobulina) en la muestra pulmonar tomada a las 12 semanas después de la infusión. Se observaron osteoblastos en el 100% de los ratones infundidos con CSM fucosiladas, en 62,5% de los infundidos con CSM, y ninguno en el grupo de salino (p=0.01). Los ratones infundidos con CSM fucosiladas presentaron un mayor número de osteoblastos osteocalcina positivos en 10 campos de gran aumento (400x) de secciones de tibia y calota con respecto a los infundidos con no fucosiladas (32 vs 5.5) (p=0.0082). Las CSM fucosiladas. Los osteoblastos humanos fueron detectados en el hueso desde la quinta a la doceava semana post-infusión. Conclusiones: Nuestro ensayo preclínico demuestra que la infusión intravenosa de CSM fucosiladas es segura y eficaz en guiar las células al hueso con un potencial más elevado de osificación en ratones NOD/SCID que las CSM no fucosiladas. Estos resultados van a permitir comenzar con un Ensayo clínico en humanos con CSM fucosiladas. ABSTRACT Objectives: Osteoporosis is a skeletal disease characterized by systemic bone loss with an increased risk to bone fractures, associated with high morbidity and mortality. Mesenchymal stem cells (MSC) from bone marrow (BM) are ideal candidates for the treatment of osteoporosis because they are able to differentiate to osteoblasts. Although bone graft of MSCs has been demonstrated, its osteotropism is low when they are administered intravenously (IV), of choice in this multifocal disease. MSC homing depends upon the adhesive interaction between MSC surface receptors and counter receptors on endothelial cells. E-selectin protein is induced on microvascular endothelium in inflammatory tissues in response to cytokines and it is constitutively expressed in BM endothelial cells. The Hematopoietic cell E and L-selectin ligand (HCELL), a glycoform of CD44, is a potent E-selectin ligand expressed constitutively in the Hematopoietic stem cells (HSC). HCELL represents the key to involve in cell recruitment to the BM. MSC express CD44 but do not express HCELL or other E-selectin ligands. Ex vivo fucosylation (adding a fucose residue) in α1-3 position of the CD44 antigen by use of the enzyme fucosyltransferase VI (FTVI) yields HCELL in MSC increasing the affinity for E-selectin and osteotropism. The aim of this preclinical study was to describe the safety and efficacy of human fucosylated MSC infused in an immunocompromised mice model (NOD/SCID). Methods: 31 NOD/SCID mice were randomized to by tail vein injection: 1x106 fucosylated MSC (n=13), 1x106 MSC (n=14) or saline (n=4). Toxicities were evaluated by a clinical score, weight and histological assessment (heart, lung, liver, spleen, kidney, gonads, brain, bone and BM). RT-PCR array-based evaluation of the expression of human β-actin and β2-microglobulin genes was performed to study biodistribution of MSC. The maintenance of genetic integrity was evaluated during in vitro culture by karyotype. Additional samples of tibia and calota were to immunostained with a polyclonal Rabbit anti-human osteocalcina to demonstrate efficacy. Osteocalcin-positive cells were identified by a dark-brown cytoplasmic precipitate. Results: There was no unexpected death and none of the mices had any acute toxicity. One mouse in the saline arm had mild chronic toxicity primarily manifested as piloerection, and moderate changes in behavior. The histology of heart, liver, kidney, spleen, gonads, brain, bone and bone marrow was normal in all mice. There were localized areas of lung inflammation in 15%, 42% and 25% of mice infused with fucosylated MSC, MSC, and saline, respectively without significance differences (p=0.28). Biodistribution was normal with except one mice (infused with MSC non-fucosylated) showed an expression of human RNA β-actin and β2-microglobulin in the lung sample taken after 12 weeks of intravenous infusion .Osteoblasts were seen in 100% of mices infused with fucosylated MSC, in 62.5%% of animals infused with MSC alone, and none in saline group (p=0.01). NOD/SCID mice infused with fucosylated MSC presented a higher number of osteoblasts positive for osteocalcin in 10 high-power fields (400x) of tibia and calvarium sections that non-fucosylated (32 vs 5.5) (p=0.0082). Human osteoblasts were detected inside the bone from the fifth to the twelfth week after infusion. Conclusions: Our preclinical trial demonstrate that the intravenous infusion of human fucosylated MSC is safe and effective in guiding the cells to bone with a higher potential to ossification in NOD/SCID mices that non-fucosylated MSC. These results are allowing to start a human clinical trial with intravenous fucosylated MSC as treatment in patients with osteoporosis

Mass spectrometry vs immunofixation for treatment monitoring in multiple myeloma

This study was supported by grants from the Centro de Inves-tigación Biomédica en Red-Área de Oncología-del Instituto de Salud Carlos III CIBERONC, CB16/12/00369, CB16/12/00400, CB16/12/00233, and CB16/12/00284, Instituto de Salud Carlos III/Subdirección General de Investigación Sanitaria FIS no. PI15/ 01956, PI15/02049, PI15/02062, PI18/01709, PI18/01673, and PI19/01451, the Cancer Research UK, FCAECC, and AIRC under the Accelerator Award Program (EDITOR).Monitoring of the monoclonal protein (M-protein) by electrophoresis and/or immunofixation (IFE) has long been used to assess treatment response in multiple myeloma (MM). However, with the use of highly effective therapies, the M-protein becomes frequently undetectable, and more sensitive methods had to be explored. We applied IFE and mass spectrometry (EXENT&FLC-MS) in serum samples from newly diagnosed MM patients enrolled in the PETHEMA/GEM2012MENOS65 obtained at baseline (n 5 223), and after induction (n 5 183), autologous stem cell transplantation (n 5 173), and consolidation (n 5 173). At baseline, the isotypes identified with both methods fully matched in 82.1% of samples; in the rest but 2 cases, EXENT&FLC-MS provided additional information to IFE with regards to the M-protein(s). Overall, the results of EXENT&FLC-MS and IFE were concordant in.80% of cases, being most discordances due to EXENT&FLC-MS but IFE cases. After consolidation, IFE was not able to discriminate 2 cohorts with different median progression-free survival (PFS), but EXENT&FLC-MS did so; furthermore, among IFE patients, EXENT&FLC-MS identified 2 groups with significantly different median PFS (P 5.0008). In conclusion, compared with IFE, EXENT&FLC-MS is more sensitive to detect the M-protein of patients with MM, both at baseline and during treatment, and provides a more accurate prediction of patients' outcome. This trial was registered at www.clinicaltrials.gov as #NCT01916252

Prognostic Value of Serum Paraprotein Response Kinetics in Patients With Newly Diagnosed Multiple Myeloma

Introduction: Response kinetics is a well-established prognostic marker in acute lymphoblastic leukemia. The situation is not clear in multiple myeloma (MM) despite having a biomarker for response monitoring (monoclonal component [MC]). Materials and Methods: We developed a mathematical model to assess the prognostic value of serum MC response kinetics during 6 induction cycles, in 373 NDMM transplanted patients treated in the GEM2012Menos65 clinical trial. The model calculated a "resistance" parameter that reflects the stagnation in the response after an initial descent. Results: Two patient subgroups were defined based on low and high resistance, that respectively captured sensitive and refractory kinetics, with progression-free survival (PFS) at 5 years of 72% and 59% (HR 0.64, 95% CI 0.44-0.93; P =.02). Resistance significantly correlated with depth of response measured after consolidation (80.9% CR and 68.4% minimal residual disease negativity in patients with sensitive vs. 31% and 20% in those with refractory kinetics). Furthermore, it modulated the impact of reaching CR after consolidation; thus, within CR patients those with refractory kinetics had significantly shorter PFS than those with sensitive kinetics (median 54 months vs. NR; P =.02). Minimal residual disease negativity abrogated this effect. Our study also questions the benefit of rapid responders compared to late responders (5-year PFS 59.7% vs. 76.5%, respectively [P <.002]). Of note, 85% of patients considered as late responders were classified as having sensitive kinetics. Conclusion: This semi-mechanistic modeling of M-component kinetics could be of great value to identify patients at risk of early treatment failure, who may benefit from early rescue intervention strategies