Summary. The urokinase‐type plasminogen activator (uPA) system, which consists of a proteinase (uPA), a receptor (uPAR or CD87) and inhibitors, is involved in proteolysis, cell migration, tissue remodelling, angiogenesis and cell adhesion. Recent findings suggest that malignant plasma cells express uPA and uPAR. The expression of these factors could represent a process by which myeloma plasma cells interact with the bone marrow (BM) environment and influence important biological events such as bone matrix degradation, plasma cell invasion and homing and, possibly, clinical evolution. We evaluated uPAR (CD87) and its soluble form (suPAR) in 49 multiple myeloma (MM) patients and correlated their expression and levels with clinico‐biological characteristics of the disease. Flow cytometric analysis demonstrated that CD87 was expressed in all MM patients. High CD87 expression was associated with higher intensity of expression of CD56 ( P = 0·038), CD38 ( P = 0·058) and CD138 ( P = 0·054) and CD45 bright positivity ( P = 0·014). suPAR levels correlated positively with soluble serum CD138 ( P = 0·001), creatinine ( P = 0·001), beta 2 ‐microglobulin ( P < 0·001), disease stage ( P = 0·017) and extra‐BM involvement ( P = 0·002). In the 46 evaluable patients, multivariate analysis showed that high levels of suPAR ( P = 0·0214) and disease stage ( P = 0·0064) were predictive of extra‐BM involvement. In multivariate Cox analysis, 13q deletion ( P = 0·0278), high soluble serum CD138 ( P = 0·0201) and high suPAR ( P = 0·0229) were the only parameters that independently affected survival. We conclude that CD87 is expressed on myeloma plasma cells and that suPAR, which predicts extra‐BM involvement and poor prognosis, possibly represents a molecule with a relevant role in the biology of MM.
Summary The in vivo effect of recombinant human erythropoietin (rHuEpo) and granulocyte colony‐stimulating factor (G‐CSF) combined treatment on CD34 + cells was evaluated by fluorescence in situ hybridization (FISH) in 13 myelodysplastic syndrome (MDS) patients with known cytogenetic abnormalities. After treatment, responsive patients presented a significantly lower proportion of FISH abnormal CD34 + cells than before treatment ( P = 0·003), and in comparison with unresponsive cases ( P = 0·007). Response to treatment was associated with a reduced degree of apoptosis in CD34 + cells ( P = 0·021): however, no difference in telomere length was observed in responsive patients after growth factor administration. Although the number of patients analysed was relatively small, the present data suggest that, in MDS patients, response to rHuEpo and G‐CSF may be related to the proliferation of karyotypically normal but potentially defective CD34 + progenitor cells.
Summary. The issue of whether, in patients affected by myelodysplastic syndromes (MDS), haematological response to cytokines, particularly to recombinant human erythropoietin (rHuEpo), is a phenomenon related to the stimulation of normal haemopoietic cells or to the differentiation of cells belonging to the abnormal clone remains an open question. To assess the pattern of response to rHuEpo treatment of bone marrow (BM) cells, we evaluated in 13 low‐risk MDS patients with known cytogenetic abnormalities the number of cytogenetically normal and abnormal cells by conventional cytogenetic analysis (CCA) and by a fluorescence in situ hybridization (FISH) technique, enabling the simultaneous visualization of FISH chromosomal abnormalities in morphologically and immunophenotypically identifiable BM elements. Patients responding to rHuEpo presented a lower number of abnormal metaphases at diagnosis in comparison with patients who did not respond (22·74% vs 76·23%, P = < 0·001). This was confirmed by the combined morphological FISH analysis, showing that, before treatment, BM samples from patients responding to rHuEpo had a lower proportion of both FISH abnormal erythroid (36·48% vs 66·93%, P = 0·002) and myeloid (40·76% vs 67·70%, P = 0·014) elements than unresponsive patients. After rHuEpo treatment, responding patients presented a significantly lower proportion of FISH abnormal erythroid precursors than observed before treatment (16·93% vs 36·48%, P = 0·017). Likewise, in responding patients, a significantly lower proportion of FISH abnormal erythroid elements (16·93% vs 66·30%, P < 0·001) was detected in comparison with unresponsive patients. These findings provide evidence that, in low‐risk MDS patients with known cytogenetic abnormalities, response to rHuEpo may be due to the proliferation of karyotypically normal erythroid precursors, possibly representing residual normal erythroid elements.
Inherited defects in the genes of blood coagulation essentially express the severity of the clinical phenotype that is directly correlated to the number of mutated alleles of the candidate leader gene (e.g., heterozygote vs. homozygote) and of possible additional coinherited traits. The F5 gene, which codes for coagulation factor V (FV), plays a two-faced role in the coagulation cascade, exhibiting both procoagulant and anticoagulant functions. Thus, defects in this gene can be predisposed to either bleeding or thrombosis. A Sanger sequence analysis detected a premature stop-codon in exon 13 of the F5 gene (c.3481C>T; p.R1161Ter) in several members of a family characterised by low circulating FV levels and contrasting clinical phenotypes. The propositus, a 29 y.o. male affected by recurrent haemorrhages, was homozygous for the F5 stop-codon and for the F5 c.1691G>A (p.R506Q; FV-Leiden) inherited from the heterozygous parents, which is suggestive of combined cis-segregation. The homozygous condition of the stop-codon completely abolished the F5 gene expression in the propositus (FV:Ag < 1%; FV:C < 1%; assessed by ELISA and PT-based one-stage clotting assay respectively), removing, in turn, any chance for FV-Leiden to act as a prothrombotic molecule. His father (57 y.o.), characterised by severe recurrent venous thromboses, underwent a complete molecular thrombophilic screening, revealing a heterozygous F2 G20210A defect, while his mother (56 y.o.), who was negative for further common coagulation defects, reported fully asymptomatic anamnesis. To dissect these conflicting phenotypes, we performed the ProC®Global (Siemens Helthineers) coagulation test aimed at assessing the global pro- and anticoagulant balance of each family member, investigating the responses to the activated protein C (APC) by means of an APC-sensitivity ratio (APC-sr). The propositus had an unexpectedly poor response to APC (APC-sr: 1.09; n.v. > 2.25), and his father and mother had an APC-sr of 1.5 and 2.0, respectively. Although ProC®Global prevalently detects the anticoagulant side of FV, the exceptionally low APC-sr of the propositus and his discordant severe–moderate haemorrhagic phenotype could suggest a residual expression of mutated FV p.506QQ through a natural readthrough or possible alternative splicing mechanisms. The coagulation pathway may be physiologically rebalanced through natural and induced strategies, and the described insights might be able to track the design of novel treatment approaches and rebalancing molecules.
Topic: 35. Quality of life and palliative care Background: Owing to advances made in the treatment of acute myeloid leukemia (AML), for example with regard to transplantation strategies, the number of AML survivors has increased over the last decades. Therefore, it becomes increasingly important to understand the long-term effects of AML therapies to better address unmet needs of these patients. Aims: An international AML survivorship project was performed by the GIMEMA in collaboration with the EORTC Leukemia and Quality of Life Group, and we herein report first results of this project by describing the prevalence of long-term health problems and comorbidities experienced by these patients, overall and by age at diagnosis. Methods: Eligibility criteria of this international study by the GIMEMA and EORTC Leukemia and Quality of Life Groups included: a diagnosis of AML (at least 20% blast in bone marrow) other than acute promyelocytic leukemia, age ≥ 18 years, at least 5 years after diagnosis, and being in AML-free status at the time of study entry. Patients were approached at the hospital or by mail and those consenting were asked to complete a Survey Booklet, which also included a comprehensive list of health problems and comorbidities derived from the validated Self-administered Comorbidity Questionnaire and health problems from other AML survivorship studies. Comorbidity burden was defined as having at least one comorbidity/health problem vs none. The comorbidity burden, as well as each comorbidity prevalence was examined overall and stratified by median age at diagnosis, identifying two groups (younger and older AML survivors). Results: Between May 2019 and February 2022, 225 AML survivors were enrolled by 24 centers across 6 countries. There were 106 (47.1%) males and 119 (52.9%) females and the median age at diagnosis was 48.3 years. Almost half of patients had an intermediate education level (i.e. up to high school) (102, 45.3%) and the majority of participants were living with a partner (187, 83.1%). Median time from AML diagnosis was 8.8 years (IQR 6.4-11.9). There were 70 survivors (31.1%) who were treated in the setting of a randomized controlled trial during primary treatment. Time from AML diagnosis, and from end of treatment to study inclusion, were significantly longer (p<0.001) in younger survivors (median age at study inclusion of 46.8 years), compared to older survivors (median age at study inclusion of 66.8 years). Similar frequencies of at least one AML relapse and allogeneic hematopoietic stem cell transplantation were observed in younger and older survivors, i.e. 21 (19.1%) vs 26 (22.6%) and 59 (53.6%) vs 63 (54.8%), respectively. One hundred ninety five survivors (86.7%) reported at least one comorbidity and 103 (45.8%) reported four or more comorbidities. The top three most prevalent comorbidities were: impaired vision (106, 47.1%), back pain (79, 35.1%), arthrosis/arthritis (76, 33.8%). The top ten most prevalent comorbidities for the overall population are depicted in Figure 1. Inspection of prevalence by age at diagnosis revealed a higher prevalence in the older survivors’ group with regard to some specific comorbidities. For example, younger survivors compared to older survivors reported a lower prevalence of hypertension (20.0% vs 41.7%), diabetes (8.2% vs 22.6%) and heart disease (1.8% vs 14.8%), respectively. Summary/Conclusion: Our findings indicate that long-term AML survivors have a substantial comorbidity burden even many years after diagnosis and may help to identify most pressing needs of these patients. Further analyses are needed to further elucidate the relationships between patient’s comorbidity and quality of life of AML survivors.Keywords: Comorbidities, Acute myeloid leukemia, Quality of life, Patient reported outcomes
Summary The child's discomfort and the cost of overnight hospitalization are clear disadvantages of prolonged esophageal pH monitoring. The aim of this study was to verify the reliability of short recording versus 24-h testing in a pediatric series with symptoms suggestive of gastro-esophageal reflux (GER) disease. A 24-h pH monitoring performed on 160 patients with either gastroenterological symptoms (n = 61), respiratory problems (n = 58), or emesis plus respiratory problems (n = 41) was reviewed. Regardless of clinical presentation, children were also classified according to age: <12 months (n = 39), 12–71 months (n = 81), and 72–168 months (n = 40). A diurnal fraction of 6 h, including at least 2 h after a meal, was compared to the entire 24-h recording in all groups with respect to the reflux index (RI) (sum of the periods with pH <3.9 expressed as percentage of time) and reflux/h. RIs of > 10% were considered positive in patients < 1 year of age, whereas RIs of >5% were considered positive in other age groups. Negative predictive values of the short recording RI ranged from 71 to 90%. Positive predictive values ranged from 50 to 83%; it was unreliable for children <12 mos (50%) and patients with emesis plus respiratory problems (64%), who were, significantly, the youngest. Reflux/h values were not in agreement for the same groups. Absence of agreement was found if the absolute value of RI was considered. In conclusion, our data show that short-term recordings may be used as an ambulatory screening test for GER in selected children, being unreliable for patients <1 year of age and for those presenting with both gastroenterological and respiratory symptoms.
