Advancing molecular imaging: a chairman’s perspective on how radiology can meet the challenge

To date, most molecular imaging techniques applied clinically have offered relatively general information about the metabolism and physiology of diseased cells and tissues. However, due to recent scientific and technological advances, much more specifically targeted molecular imaging probes (e.g., reporter gene probes, whole cell-tracking probes, and probes for localizing specific biomolecules) are now being used in preclinical research and, in some cases, translated to the clinical setting. As a result, the imaging community is poised to help lead a revolution in personalized, molecularly targeted medicine. This article considers the importance of molecular imaging for advancing research and clinical care both within individual institutions and across the medical field. It outlines specific steps that leaders in academic radiology can take to hasten progress in molecular imaging and explains why they must have the courage to reach across traditional interdisciplinary boundaries and advocate for major investments in equipment, education, and personnel.     

Clinical applications in molecular imaging

Molecular imaging is aimed at the noninvasive in vivo characterization and measurement of processes at a cellular and molecular level with clinical imaging methods. Contrast agents are constructed to target markers that are specific either for certain diseases or for functional states of specialized tissues. Efforts are currently focused mainly on processes involved in angiogenesis, inflammation, and apoptosis. Cell tracking is performed for diagnostic purposes as well as for monitoring of novel cell therapies. Visualization of these processes would provide more precise information about disease expansion as well as treatment response, and could lead to a more individualized therapy for patients. Many attempts have shown promising results in preclinical studies; however, translation into the clinic remains a challenge. This applies especially to paediatrics because of more stringent safety concerns and the low prevalence of individual diseases. The most promising modalities for clinical translation are nuclear medicine methods (positron emission tomography [PET] and single photon emission CT [SPECT]) due to their high sensitivity, which allows concentrations below biological activity. However, special dose consideration is required for any application of ionizing radiation especially in children. While very little has been published on molecular imaging in a paediatric patient population beyond fluorodeoxyglucose (FDG)-PET and metaiodobenzylguanidine (MIBG) tracers, this review will attempt to discuss approaches that we believe have promise for paediatric imaging. These will include agents that already reached clinical trials as well as preclinical developments with high potential for clinical application.     

Aktualne zasady diagnostyki oraz zmiany w klasyfikacji wrodzonej łamliwości kości (Osteogenesis imperfecta)

The definition and classification of Osteogenesis imperfecta (OI) have been transformed over the last two decades. In the past, it has been described as a type I collagenopathy due to identified mutations in COL1A1 and COL1A2 genes encoding collagen type I which led to all OI cases. At least 10 genes involved in the process of type I collagen biosynthesis have been described until now. COL1A1/2 genes are responsible for almost 90% of OI prevalence. However, there is a number of newly discovered genes which encode proteins involved in posttranslational modification of procollagen, whereas mutations in these genes lead to approximately 10% types of OI inherited recessively. In this review, we provide an update on the traditional Sillence classification of the disease and we also report recent developments in understanding of underlying mechanisms responsible for OI. The paper discusses currently published modification of the OI classification (new types of the disease: V–XV) and a new approach to the diagnostic methods focusing mainly on phenotypic manifestation, independent of the genetic or molecular background. We point out the importance of clinical symptoms and severity of the disorder of which both appear the most useful criteria for management of OI patients in everyday practice.     

Just taller or more bone? The impact of growth hormone on osteogenesis imperfecta and idiopathic juvenile osteoporosis

There are scant data to date on the use of GH therapy in osteogenesis imperfecta (OI) or in idiopathic juvenile osteoporosis (IJO). In OI, we review three clinical trials (aggregate study population 46), and 13 patient reports from the NCGS, and two independent patient reports. Based on evidence of increased growth rate versus some reported increased fracture rate, we conclude that GH should probably not be used as first-line therapy in OI, pending further data from clinical trials. There are no published reports of the use of GH therapy in IJO and only one patient with documented IJO enrolled in the NCGS. Since IJO presents in the immediate prepubertal period and appears to improve naturally during puberty, it is difficult to differentiate the effects of GH therapy from those of puberty; therefore, we conclude that GH in IJO should currently be used only for research and not in clinical practice.     

COL1A2新突变致胎儿严重成骨不全症1例报告

目的 总结1例COL1A2新突变致胎儿严重成骨不全症（OI）的临床特征及基因突变的特点,为胎儿产前咨询提供依据。方法 对产检B超检查示OI可能的胎儿流产组织抽提DNA进行基因型分析,自行设计COL1A1和COL1A2所有外显子及剪接区域的引物。利用Sanger测序法对胎儿行COL1A1和COL1A2 基因外显子及剪接区域的测序分析并行父母验证。依据人类基因突变数据库（HGMD）专业版,对COL1A2突变所致疾病临床表型行文献复习。结果 胎儿的COL1A1和COL1A2基因均检测出变异位点。COL1A2基因检测到杂合突变（c.3142G>T, p. Glu1048Cys）在寡核苷酸多态性数据库、HGMD及Ⅰ型胶原蛋白突变数据库均未见报道,结合胎儿父母验证为新发突变,对比公共数据库及在线预测软件预测该突变类型为致病突变。在HGMD专业版中搜索COL1A2,共检索到387个COL1A2致病突变,与21种疾病及其亚型相关。92%的突变引起OI或其亚型,还可引起Ehlers-Danlos综合征或其亚型。结合COL1A2突变所致疾病临床表型行文献复习,本文报告胎儿符合Ⅱ型OI。结论 产前通过超声影像结合基因分型诊断胎儿为COL1A2基因新发突变（c.3142G>T, p. Glu1048Cys）所致Ⅱ型OI; COL1A2基因编码蛋白长链双螺旋的400～480氨基酸及MLBR 3区域中甘氨酸被天冬氨酸或谷氨酸替代,多导致严重表型的OI;本文为产前准确预测胎儿结局、指导临床决策提供依据。     

What is new in genetics and osteogenesis imperfecta classification?

ObjectiveLiterature review of new genes related to osteogenesis imperfecta (OI) and update of its classification.SourcesLiterature review in the PubMed and OMIM databases, followed by selection of relevant references.Summary of the findingsIn 1979, Sillence et al. developed a classification of OI subtypes based on clinical features and disease severity: OI type I, mild, common, with blue sclera; OI type II, perinatal lethal form; OI type III, severe and progressively deforming, with normal sclera; and OI type IV, moderate severity with normal sclera. Approximately 90% of individuals with OI are heterozygous for mutations in the COL1A1 and COL1A2 genes, with dominant pattern of inheritance or sporadic mutations. After 2006, mutations were identified in the CRTAP, FKBP10, LEPRE1, PLOD2, PPIB, SERPINF1, SERPINH1, SP7, WNT1, BMP1, and TMEM38B genes, associated with recessive OI and mutation in the IFITM5 gene associated with dominant OI. Mutations in PLS3 were recently identified in families with osteoporosis and fractures, with X-linked inheritance pattern. In addition to the genetic complexity of the molecular basis of OI, extensive phenotypic variability resulting from individual loci has also been documented.ConclusionsConsidering the discovery of new genes and limited genotype-phenotype correlation, the use of next-generation sequencing tools has become useful in molecular studies of OI cases. The recommendation of the Nosology Group of the International Society of Skeletal Dysplasias is to maintain the classification of Sillence as the prototypical form, universally accepted to classify the degree of severity in OI, while maintaining it free from direct molecular reference.     

Extracranial outflow of particles solved in cerebrospinal fluid: Fluorescein injection study

Cerebrospinal fluid is thought to be mainly absorbed into arachnoid granules in the subarachnoid space and drained into the sagittal sinus. However, some observations such as late outbreak of arachnoid granules in fetus brain and recent cerebrospinal fluid movements study by magnetic resonance images, conflict with this hypothesis. In this study, we investigated the movement of cerebrospinal fluid in fetuses. Several kinds of fluorescent probes with different molecular weights were injected into the lateral ventricle or subarachnoid space in mouse fetuses at a gestational age of 13 days. The movements of the probes were monitored by live imaging under fluorescent microscope. Following intraventricular injection, the probes dispersed into the 3rd ventricle and aqueduct immediately, but did not move into the 4th ventricle and spinal canal. After injection of low and high molecular weight conjugated probes, both probes dispersed into the brain but only the low molecular weight probe dispersed into the whole body. Following intra‐subarachnoid injection, both probes diffused into the spinal canal gradually. Neither probe dispersed into the brain and body. The probe injected into the lateral ventricle moved into the spinal central canal by the fetus head compression, and returned into the aqueduct by its release. We conclude this study as follows: (i) The movement of metabolites in cerebrospinal fluid in the ventricles will be restricted by molecular weight; (ii) Cerebrospinal fluid in the ventricle and in the subarachnoid space move differently; and (iii) Cerebrospinal fluid may not appear to circulate. In the event of high intracranial pressure, the fluid may move into the spinal canal.     

Osteogenesis imperfecta: contribution to pathobiochemistry

Osteogenesis imperfecta (OI) is a hereditary disease characterized by increased bone fragility and marked skeletal deformities. As a generalized connective tissue disorder, many patients present with other typical symptoms, such as blue sclerae, dentinogenesis imperfecta, impaired hearing, joint laxity, and easy bruising of the skin. According to clinical and genetic characteristics, Sillence classified four different groups. Metabolic alterations of connective tissue components are thought to be responsible for the pathogenesis of OI. Collagen, the main constituent of connective tissue, was analyzed in autoptic tissue and/or skin fibroblasts from patients with OI. In fibroblast culture of patients with OI group I, the range of synthesized collagen type III is elevated to 15-48% (normal up to 15%). Patients in groups II and III show an increased presence of hydroxylysine in alpha-chains of collagen types I and III. The hydroxylation of lysyl residues of the cartilage specific collagen type II is slightly elevated. In both groups, the hydroxypyroline content in all tested collagen types was normal. In our investigation, the collagen of patients with OI group IV appeared normal. Although the clinical features of patients with OI of all groups were not homogeneous, OI group III and some subtypes of group II had similar clinical courses and biochemical findings. In addition, there are patients with OI who present with clinical symptoms, but who cannot be classified into any of the known groups. For a better differentiation, biochemical examination of collagen should be performed complementary to clinical and genetic criteria.     

Schwerwiegende Netzhauterkrankungen im Kindesalter

This paper summarizes current diagnostic and therapeutic methods for selected paediatric retinal diseases which already today or in the near future will have a significant impact on the prognosis. New aspects of retinopathy of prematurity (ROP), hereditary global or central retinal dystrophies with childhood manifestation as well as retinal disorders associated with infantile nystagmus syndrome (INS) will be presented. Important diagnostic methods are full-field ERG, multifocal ERG (mfERG), fundus controlled perimetry, optical coherence tomography (OCT), fundus autofluorescence (FAF) and wide-field digital imaging (WFDI). In addition to the clinical examination these methods allow gene directed molecular diagnostics. An exact classification is essential for precise counseling of the families as to prognosis and recurrence risk of the disease. Early stages of organic disorders can thus be safely differentiated from functional visual loss and primary pathologies of the optic nerve or central visual pathways.     

胎粪吸入性肺炎应用常频与高频通气的研究

目的　 评估常频通气和高频通气治疗胎粪吸入性肺炎 (MAS)所致呼吸衰竭的临床与实验对比疗效。 方法 　实验方面 :健康白兔 ,气管注入胎粪混悬液 ( 3～ 5ml/kg) ,呼吸衰竭模型出现后随机分组 :①继续常频通气 (CMV) 3h后转为高频通气 (HFV) 3h(CMV HFV组 ) ;②转为HFV 3h后再转为CMV 3h(HFV CMV组 )。 2 0例MAS患儿入院后即用CMV治疗 ,当所用吸入氧浓度 (FiO2 ) >0 82h左右 ,经皮氧饱和度 (TcPO2 )仍 <90 %,改为HFV治疗 ;当FiO2 >0 82h左右 ,TcPO2 能维持在90 %以上 ,继续用CMV治疗。 10例继续应用CMV治疗 ,10例改用HFV治疗。 结果 　动物出现呼吸衰竭模型时 ( 0h)的氧合指数 (OI)二组无差异 ,CMV HFV组在治疗后 3hOI无明显变化 ,至 6hOI有上升 ;HFV CMV组治疗后 3hOI较 0h时有下降 ,两组动物治疗后 3h的OI的组间比较有差异。 2 0例患儿中 ,CMV组治愈 7例 ,死亡 2例 ,放弃 1例 ;HFV组治愈 7例 ,放弃 2例 ,死亡1例。HFV组患儿治疗后 1hOI较 0h有下降 ,至治疗后 6h ,OI与 0h比较差异显著 ;CMV组治疗后 1hOI有明显下降 ,但至治疗后 6h下降幅度不明显。二组患儿的呼吸机应用时间和用氧时间均无差异。 结论 　MAS用HFV治疗后的氧合作用较CMV改善明显和吸入氧浓度下降迅速     

Molecular ultrasound imaging and its potential for paediatric radiology

US is a low-cost, real-time imaging modality that is the most used diagnostic tool in paediatric radiology. Reasons include the improved US image quality in children as compared to adults and the demand for avoiding X-rays as much as possible because children are more sensitive to radiation than adults. Stabilized microbubbles have been approved as US contrast agents for adults and show great potential in improving the diagnostic accuracy for many diseases. Initial studies show that in paediatric radiology contrast-enhanced US could also be beneficial for more than just the diagnosis of vesicoureteral reflux, if US contrast agents were approved for children. Molecular US imaging utilizes microbubbles conjugated to biomolecules that target intravascular disease-specific molecules. Many preclinical studies show that molecular US imaging is a highly sensitive tool to detect neovascularisation, inflammation and cardiovascular diseases. Its main advantages are the higher informative value, the longer persistence of the label at the target lesion and the chance to work with lower contrast agent dosages. Now, clinical translation of molecular US appears at the horizon. This review article reports on the current status of molecular US imaging and discusses its potential for paediatric radiology.     

PET imaging in pediatric neuroradiology: current and future applications

Molecular imaging with positron emitting tomography (PET) is widely accepted as an essential part of the diagnosis and evaluation of neoplastic and non-neoplastic disease processes. PET has expanded its role from the research domain into clinical application for oncology, cardiology and neuropsychiatry. More recently, PET is being used as a clinical molecular imaging tool in pediatric neuroimaging. PET is considered an accurate and noninvasive method to study brain activity and to understand pediatric neurological disease processes. In this review, specific examples of the clinical use of PET are given with respect to pediatric neuroimaging. The current use of co-registration of PET with MR imaging is exemplified in regard to pediatric epilepsy. The current use of PET/CT in the evaluation of head and neck lymphoma and pediatric brain tumors is also reviewed. Emerging technologies including PET/MRI and neuroreceptor imaging are discussed.     

Martin-Bell syndrome. Improved possibilities for molecular genetic diagnosis]

In a former molecular segregation analysis of fra x mental retardation in 27 families at risk we had used marker gene probes with a relatively high recombination fraction. Thus, the resulting risk for a false diagnosis was comparatively high. To diminish this risk, all families were reanalyzed with the newly invented and more closely linked gene probes RN1A, VK23B, VK21C and U6.2. Using these probes as molecular markers we performed Southern hybridization experiments. The remaining diagnostic risk due to recombination events could be reduced to 2% up to 20% compared to preanalysis. The portion of informative families (91%) is in good agreement with the expected cumulative heterozygosis frequency of 93.4% for all 4 markers investigated. This high frequency and the very low remaining risk for a false diagnosis therefore enable a far more precise molecular diagnostic of the Martin-Bell-syndrome.     

Precision medicine in pediatric oncology: Lessons learned and next steps

The maturation of genomic technologies has enabled new discoveries in disease pathogenesis as well as new approaches to patient care. In pediatric oncology, patients may now receive individualized genomic analysis to identify molecular aberrations of relevance for diagnosis and/or treatment. In this context, several recent clinical studies have begun to explore the feasibility and utility of genomics‐driven precision medicine. Here, we review the major developments in this field, discuss current limitations, and explore aspects of the clinical implementation of precision medicine, which lack consensus. Lastly, we discuss ongoing scientific efforts in this arena, which may yield future clinical applications.     

Maternal adjustment,parenting and child behaviour in families of school-aged twins conceived after IVF and ovulation induction

BACKGROUND: Previous studies that have examined the long-term effects of infertility and assisted reproductive technology on parenting and child behaviour in families with twins have suffered from methodological problems. This study compared measures of parental adjustment, parenting and child behaviour in families with 5-year-old twins who were conceived after in vitro fertilisation (IVF) or ovulation induction (OI) with families whose twins were naturally conceived (NC). METHODS: The families who conceived via IVF/OI (N = 121) were identified from an epidemiological study of twins and matched to families who were conceived naturally (N = 121) on the basis of eleven child and family variables. Mothers were interviewed in their homes for the study. RESULTS: No significant differences were observed between the IVF/OI families and the NC families on measures of parental adjustment or parent and teacher ratings of the twins' behaviour. IVF/OI mothers and their partners agreed with each other about discipline more than NC couples, but otherwise no other differences in parenting were found. CONCLUSIONS: Overall, this study provides evidence that families who conceive twins following IVF/OI are functioning well and that the experience of fertility treatment does not lead to long-term difficulties for parents or children.     

Longitudinal study of 94 symptomatic infants with perinatally acquired human immunodeficiency virus infection. Evidence for a bimodal expression of clinical and biological symptoms

To better define the clinical and biological evolution of infants after vertical human immunodeficiency virus type 1 infection, we analyzed 94 consecutive infected patients followed up after their first clinical symptoms. The expression of clinical symptoms and biological abnormalities followed a bimodal distribution, some patients having an early and severe disease and the others having a slowly progressive one. One third of our patients suffered from early onset of opportunistic infection (OI). These patients had a significantly higher incidence of severe encephalopathy compared with patients without OI. The rate of survival at 3 years was 48% +/- 24%. In contrast, the patients without early OI or severe encephalopathy had a probability of survival at 3 years of 97% +/- 3%. This probability was not modified by the occurrence of bacterial infection or lymphoid interstitial pneumonitis. Lymphoid interstitial pneumonitis occurred at a mean age of 29 months, significantly later than OI or severe encephalopathy. Laboratory results at initial examination were correlated with clinical symptoms. Thus, when the number of CD4 lymphocytes was less than 500/mm3, children suffered more frequently from life-threatening symptoms (OI and severe encephalopathy): 15 of 22 vs 14 of 69. The same was true when the lymphocytes did not proliferate after antigenic stimulation, when anti-p18 and/or anti-p25 antibodies were absent in the serum, and when p24 antigen was detected in serum. Finally, severe encephalopathy was associated with low anti-human immunodeficiency virus cerebrospinal fluid antibody titer, whereas 88% of patients with moderate or no encephalopathy had signs of intrathecal anti-human immunodeficiency virus antibody synthesis. In conclusion, a subgroup of patients expressed very early signs of severe immunodeficiency and encephalopathy, whereas the majority of patients had a longer survival and less severe clinical symptoms during their first years of life than previously thought.     

Osteogenesis imperfecta and its molecular diagnosis by determination of mutations of type I collagen genes

Osteogenesis Imperfecta is a genetic disorder of increased bone fragility and low bone mass. Most cases are caused by a mutation in one of the two genes coding for the type I collagen protein. The correct clinical diagnosis of OI can be difficult sometimes, because of the wide phenotypic range. Therefore collagen I genes mutation identification can be helpful. We screened 23 patients by direct sequencing of the exons encoding the collagen protein. We identified 18 different mutations, while 5 cases were negative because of an uncertain clinical diagnosis or an atypical form of OI not related to collagen I genes. The current medical and pharmaceutical treatments are only symptomatic and do not alter the course of collagen mutations. Cells and gene therapies as potential treatments for OI have therefore to be actively investigated.     

Recent progress in diagnosis and treatment of osteogenesis imperfecta

Osteogenesis imperfecta (OI) is an inheritable disorder characterized by bone fragility with various symptoms of connective tissue disorders. OI is commonly classified by Sillence's classification into four types according to the clinical features. The cardinal symptom is pathologic fracture, which is often recognized before birth, is frequent during infancy and childhood, then decreases at puberty. Bone mineral density is markedly decreased in OI, especially of the lumbar spine. Bone deformities are frequently observed in the long bones of the extremities, and spinal deformities and compression fractures are also common. Growth retardation is extremely severe, especially in type III. Calcitonin has been the most common therapy for OI. Recently, bisphosphonates have been found to be potent drugs that increase bone mass in OI patients. To prevent further fracture or bone deformity, appropriate orthopedic managements, including intramedullary rodding, are critically important. Growth hormone is effective in stimulating bone growth during childhood. The pathogenesis of OI is quantitative or qualitative abnormalities of type I collagen. The clinical features of each type usually correspond to the type of mutation. Several possibilities for gene therapy have been proposed.     

Cardiomyocyte death: insights from molecular and microstructural magnetic resonance imaging

Cardiomyocytes can die via necrosis, apoptosis, and autophagy. Although the molecular signals and pathways underlying these processes have been well elucidated, the pathophysiology of cardiomyocyte death remains incompletely understood. This review describes the development and application of novel imaging techniques to detect and characterize cardiomyocyte death noninvasively in vivo. It focuses on molecular and microstructural magnetic resonance images (MRIs) and their respective abilities to image cellular events such as apoptosis, inflammation, and myofiber architecture. These in vivo imaging techniques have the potential to provide novel insights into the mechanisms of cardiomyocyte death and to help guide the development of novel cardioprotective therapies.     

Parental mosaicism and autosomal dominant mutations causing structural abnormalities of collagen I are frequent in families with osteogenesis imperfecta type III/IV

Abstract Protein-chemical and molecular studies were conducted on all osteogenesis imperfecta (OI) type III/IV patients referred to our hospital during the last 15 y. Of a total of 16 OI type III/IV patients studied, 15 patients were heterozygous for a mutation in one of the two genes coding for collagen I, COL1A1 or COL1A2. Cultured fibroblasts from these 15 patients produced both normal and abnormal collagen I molecules, pointing to a dominant-negative effect of the mutation. Nine mutations had not been described previously. Parental mosaicism was demonstrated in three families. In the 16th child the causative mutation was not found. In conclusion, OI type III/IV in most patients of Western European ancestry is caused by dominant mutations in the genes for collagen I, and recurrence of OI is caused in most cases by parental gonadal mosaicism.