(1)
The high-grade neuroepithelial tumor of the central nervous system with BCOR alteration (HGNET-BCOR) is a highly malignant tumor. Preclinical models and molecular targets are urgently required ...for this cancer. Previous data suggest a potential role of insulin-like growth factor (IGF) signaling in HGNET-BCOR. (2)
The primary HGNET-BCOR cells PhKh1 were characterized by western blot, copy number variation, and methylation analysis and by electron microscopy. The expression of
and
was assessed by qRT-PCR. The effect of chemotherapeutics and IGF1R inhibitors on PhKh1 proliferation was tested. The phosphorylation of IGF1R and downstream molecules was assessed by western blot. (3)
Phkh1 cells showed a DNA methylation profile compatible with the DNA methylation class "HGNET-BCOR" and morphologic features of cellular cannibalism.
and
were highly expressed by three HGNET-BCOR tumor samples and PhKh1 cells. PhKh1 cells were particularly sensitive to vincristine, vinblastine, actinomycin D (IC
< 10 nM for all drugs), and ceritinib (IC
= 310 nM). Ceritinib was able to abrogate the proliferation of PhKh1 cells and blocked the phosphorylation of IGF1R and AKT. (4)
IGF1R is as an attractive target for the development of new therapy protocols for HGNET-BCOR patients, which may include ceritinib and vinblastine.
Papillary renal cell carcinoma (PRCC) is a rare entity in children with no established therapy protocols for advanced diseases. Immunotherapy is emerging as an important therapeutic tool for ...childhood cancer. Tumor cells can be recognized and killed by conventional and unconventional T cells. Unconventional T cells are able to recognize lipid antigens presented via CD1 molecules independently from major histocompatibility complex, which offers new alternatives for cancer immunotherapies. The nature of those lipids is largely unknown and α-galactosylceramide is currently used as a synthetic model antigen. In this work, we analyzed infiltrating lymphocytes of two pediatric PRCCs using flow cytometry, immunohistochemistry and qRT-PCR. Moreover, we analyzed the CD1d expression within both tumors. Tumor lipids of PRCC samples and three normal kidney samples were fractionated and the recognition of tumor own lipid fractions by unconventional T cells was analyzed in an
assay. We identified infiltrating lymphocytes including γδ T cells and iNKT cells, as well as CD1d expression in both samples. One lipid fraction, containing ceramides and monoacylglycerides amongst others, was able to induce the proliferation of iNKT cells isolated from peripheral blood mononuclear cells (PBMCs) of healthy donors and of one matched PRCC patient. Furthermore, CD1d tetramer stainings revealed that a subset of iNKT cells is able to bind lipids being present in fraction 2 via CD1d. We conclude that PRCCs are infiltrated by conventional and unconventional T cells and express CD1d. Moreover, certain lipids, present in pediatric PRCC, are able to stimulate unconventional T cells. Manipulating these lipids and T cells may open new strategies for therapy of pediatric PRCCs.
The insulin-like growth factor (IGF) pathway plays an important role in several brain tumor entities. However, the lack of inhibitors crossing the blood-brain barrier remains a significant obstacle ...for clinical translation. Here, we targeted the IGF pathway using ceritinib, an off-target inhibitor of the IGF1 receptor (IGF1R) and insulin receptor (INSR), in a pediatric patient with an unclassified brain tumor and a
(
) germline mutation. Pathway analysis of the tumor revealed activation of the sonic hedgehog (SHH), the wingless and integrated-1 (WNT), the IGF, and the Notch pathway. The proliferation of the patient tumor cells (225ZL) was inhibited by arsenic trioxide (ATO), which is an inhibitor of the SHH pathway, by linsitinib, which is an inhibitor of IGF1R and INSR, and by ceritinib. 225ZL expressed INSR but not IGF1R at the protein level, and ceritinib blocked the phosphorylation of INSR. Our first personalized treatment included ATO, but because of side effects, we switched to ceritinib. After 46 days, we achieved a concentration of 1.70 µM of ceritinib in the plasma, and after 58 days, MRI confirmed that there was a response to the treatment. Ceritinib accumulated in the tumor at a concentration of 2.72 µM. Our data suggest ceritinib as a promising drug for the treatment of IGF-driven brain tumors.
Osteosarcoma (OS) is the second most common cause of cancer-related death in pediatric patients. The insulin-like growth factor (IGF) pathway plays a relevant role in the biology of OS but no IGF ...targeted therapies have been successful as monotherapy so far. Here, we tested the effect of three IGF specific inhibitors and tested ceritinib as an off-target inhibitor, alone or in combination with dasatinib, on the proliferation of seven primary OS cells. Picropodophyllin, particularly in combination with dasatinib and the combination ceritinib/dasatinib were effective in abrogating the proliferation. The ceritinib/dasatinib combination was applied to the primary cells of a 16-year-old girl with a long history of lung metastases, and was more effective than cabozantinib and olaparib. Therefore, the combination was used to treat the patient. The treatment was well tolerated, with toxicity limited to skin rush and diarrhea. A histopathological evaluation of the tumor after three months of therapy indicated regions of high necrosis and extensive infiltration of macrophages. The extension of the necrosis was proportional to the concentration of dasatinib and ceritinib in the area, as analysed by an ultra performance liquid chromatography-tandem mass spectrometer (UPLC-MS/MS). After the cessation of the therapy, radiological analysis indicated a massive growth of the patient's liver metastases. In conclusion, these data indicate that the combination of ceritinib/dasatinib is safe and may be used to develop new therapy protocols.
Abstract
The treatment of relapsed pediatric cancers represents a challenge, also due to possible underlying rare germline mutations. The introduction of next sequencing technologies allows a deeper ...understanding of molecular mechanisms involved in cancer progression helping in the identification of personalized therapies for these patients. However, most tumors harbor several pathway alterations and it is necessary to prioritize the treatment options. Here we applied the concept of personalized therapy to a pediatric patient with an anaplastic ependymoma and who relapsed three times under standard chemio and radiotherapy. By RNA sequencing we identified in the tumor a Notch1 mutation which was confirmed by Sanger Sequencing and which was also present in the blood DNA. Pathway analysis revealed an activation of the sonic hedgehog (SHH), the WNT, the IGF and the Notch pathway. In an attempt to prioritize possible targeted therapies, we treated the patient-specific tumor cells with relevant inhibitors in vitro. The cells were particularly sensitive to Arsenic Trioxide, an inhibitor of the SHH pathway, and to ceritinib, an off-target inhibitor of IGF-1R. Combination of Arsenic Trioxide with chemotherapy reduced the growth rate of the tumor. However, because we were not able to reach the needed Arsenic concentration in CSF, we switched to ceritinib. The patient is currently receiving the first cycle of ceritinib and chemotherapy. In the era of personalized medicine, the molecular analysis of the tumor and the use of patient-specific tumor cells may represent a powerful tool to identify new therapeutic treatments for patients with underlying rare mutations.
High-grade neuroepithelial tumor of the central nervous system with BCOR alteration (HGNET-BCOR) is a rare, highly malignant tumor. At the time of this publication, no standard protocol exists to ...treat this tumor entity. In this work, we tested the responsiveness of the primary culture PhKh1 derived from tumor tissue from a pediatric HGNET-BCOR patient (P1) to inhibitors of the Sonic hedgehog pathway combined with radiation. The SMO inhibitors vismodegib and itraconazole had low effect on the proliferation of the PhKh1 cells. However, the GLI inhibitor arsenic trioxide reduced the expression of GLI target genes in the PhKh1 cells and in combination with radiotherapy significantly decreased their clonogenic potential. PhKh1 cells resistant to arsenic trioxide were characterized by the overexpression of molecular chaperones. We combined arsenic trioxide and radiation in the relapse therapy protocol of P1, achieving complete remission after seven weeks. Clinical remission lasted for six months, when P1 developed systemic metastases. Meanwhile, an increase in the concentration of circulating tumor DNA carrying a BCOR internal tandem duplication was observed. Molecular characterization of a second patient (P2) was also performed. In P2, we detected a larger tandem duplication and greater activation of the Sonic hedgehog pathway than in P1. These findings suggest that combining arsenic trioxide with radiotherapy may represent a new therapeutic approach. Moreover, peripheral blood analysis for circulating tumor DNA could help in the early detection of systemic metastases.
High grade neuroepithelial tumor of the central nervous system with BCOR alteration (CNS HGNET-BCOR) is a recently described new tumor entity which was formerly diagnosed with diverse histological ...diagnoses, for example as ependymoma. This tumor predominantly affects children and has a dismal prognosis. No standard therapies for this entity exist so far. Recently we described the activation of the Sonic hedgehog (SHH) and the WNT signaling pathway in this tumor and described a primary cell culture (PhKh1) isolated from a skull metastasis of a seven years old patient. We also detected a high expression of IGF2, which is known to be required for SHH signaling in medulloblastoma. IGF2 signals through IGF1R leading to cell growth via the downstream PI3K/Akt pathway. In pediatric brain tumors high expression of IGF2 has been described in ependymoma and may acts as a molecular target. Here we analyzed the expression of IGF2 in our transcriptome data of 12 pediatric brain tumors patients (6 medulloblastoma, 2 glioblastoma, 2 anaplastic astrocytoma, 1 HGNET-BCOR and 1 ependymoma relapse). The highest expression of IGF2 was detected in the HGNET-BCOR and ependymoma samples. The high expression of IGF2 was maintained in the PhKh1 cell line and was similar to the expression detected in a primary cell culture isolated from the ependymoma relapse. Incubation with increasing concentrations of an antibody that blocks IGF1R resulted in a great reduction of the cell proliferation of the PhKh1 cell line but not of an IGF2-negative control cell line. In conclusion, the high expression of IGF2 may be a common feature of HGNET-BCOR and ependymoma and may represent a target for new approaches. Several monoclonal antibodies and TKIs for IGF1R are being tested in preclinical and early phase clinical studies and may become relevant in the management of this new and aggressive tumor entity.
Cytotoxic T lymphocytes are effector CD8
T cells that eradicate infected and malignant cells. Here we show that the transcription factor NFATc1 controls the cytotoxicity of mouse cytotoxic T ...lymphocytes. Activation of Nfatc1
cytotoxic T lymphocytes showed a defective cytoskeleton organization and recruitment of cytosolic organelles to immunological synapses. These cells have reduced cytotoxicity against tumor cells, and mice with NFATc1-deficient T cells are defective in controlling Listeria infection. Transcriptome analysis shows diminished RNA levels of numerous genes in Nfatc1
CD8
T cells, including Tbx21, Gzmb and genes encoding cytokines and chemokines, and genes controlling glycolysis. Nfatc1
, but not Nfatc2
CD8
T cells have an impaired metabolic switch to glycolysis, which can be restored by IL-2. Genome-wide ChIP-seq shows that NFATc1 binds many genes that control cytotoxic T lymphocyte activity. Together these data indicate that NFATc1 is an important regulator of cytotoxic T lymphocyte effector functions.NFAT nuclear translocation has been shown to be required for CD8
T cell cytokine production in response to viral infection. Here the authors show NFATc1 controls the cytotoxicity and metabolic switching of activated CD8
T cells required for optimal response to bacteria and tumor cells.
Long-term effects on infants of mothers with SARS-CoV-2 infection during pregnancy are increasingly discussed in the literature. Besides potential neurodevelopment impairments after intrauterine ...SARS-CoV-2 exposure, there might be differences in the postnatal pediatric care of those children, like the timing of preventive medical examinations (PME) or vaccinations. In this first national long-term follow-up study of women included in the CRONOS registry, we describe maternal impressions of their child´s development and the prevalence of regulatory disorders, and we analyze the timing of PMEs and vaccinations.
773 women who were enrolled between April 3, 2020, and August 24, 2021, in the CRONOS registry were eligible to be contacted by the study coordinators and asked to fill out a web-based questionnaire.
110/773 (14%) women completed the questionnaire. Their children were between the ages of 12 and 31 months (median 20 months). All mothers were satisfied with their child´s development, milestones were achieved in a timely fashion. The reported prevalence for excessive crying, sleeping, and feeding disorders was 11%, 18-32%, and 7%, respectively. PMEs were mostly not delayed, but only 54% of infants received their first vaccination within their first 60 days of life.
In summary, our exploratory findings suggest that developmental milestones in infancy are reached in time after maternal SARS-CoV-2 infection during pregnancy. However, there are effects on the implementation of PMEs and vaccinations. EINFüHRUNG: In der Literatur werden zunehmend potenzielle Langzeitfolgen für Säuglinge nach intrauteriner SARS-CoV-2-Exposition diskutiert. Neben möglichen Beeinträchtigungen der neurologischen Entwicklung können Unterschiede in der pädiatrischen postnatalen Betreuung bei diesen Kindern z. B. bei der Durchführung von Vorsorgeuntersuchungen (sog. U´s) oder Impfungen bestehen. In dieser ersten nationalen Langzeit-Follow-up-Studie aus dem CRONOS-Register beschreiben wir mütterliche Eindrücke zur Entwicklung ihres Kindes, sowie die Prävalenz von Regulationsstörungen. Wir analysieren den Zeitpunkt von U´s und Impfungen.
773 Frauen, die zwischen dem 03.04.2020 und dem 24.08.2021 in CRONOS aufgenommen wurden, wurden von den Studienkoordinatoren kontaktiert und gebeten, einen webbasierten Fragebogen auszufüllen.
110/773 (14%) Frauen füllten den Fragebogen aus, ihre Kinder waren zwischen 12 und 31 Monate alt (Median 20 Monate). Alle Mütter waren mit der Entwicklung ihres Kindes zufrieden, Meilensteine der Entwicklung wurden zeitgerecht erreicht. Die berichtete Prävalenz für exzessives Schreien, Schlaf- und Fütterstörungen betrug 11%, 18-32% bzw. 7%. U´s wurden meist zeitgerecht durchgeführt, aber nur 54% der Säuglinge erhielten ihre erste Impfung innerhalb der ersten 60 Lebenstage.
Zusammenfassend deuten unsere explorativen Ergebnisse darauf hin, dass Entwicklungsmeilensteine im Säuglingsalter nach mütterlicher SARS-CoV-2-Infektion in der Schwangerschaft rechtzeitig erreicht werden. Es zeigen sich jedoch Auswirkungen auf die Durchführung von Vorsorgen und Impfungen.
Pregnant women are at an increased risk of severe COVID-19 and adverse pregnancy outcomes; data on maternal long-term outcome is scarce. We analyzed long-term follow-ups on women who experienced a ...SARS-CoV-2 infection during pregnancy to evaluate post-COVID symptoms, particularly fatigue, and their association with quality of life (QoL).
773 women who enrolled in the CRONOS registry between April 2020 and August 2021 were contacted for follow-up from December 2022 to April 2023. Data was gathered through a web-based questionnaire. Subsequently, study coordinators matched the follow-up data with the existing CRONOS data.
110/773 (14%) women provided data. 20.9% experienced only acute symptoms during their SARS-CoV-2 infection in pregnancy, while 2.7% women experienced symptoms lasting longer than 4 weeks (long COVID). Symptoms lasting longer than 12 weeks (post-COVID) were reported by 63.6% women and occurred more often after severe COVID-19. Fatigue was the most frequently reported symptom (88%), with 55% of women still experiencing it more than one year after initial infection. 76% of women rated their QoL as "good" or "very good". Women experiencing post-COVID reported a significantly lower QoL.
This is the first German long-term data on women after SARS-CoV-2 infection during pregnancy, showing a high rate of post-COVID, a persistence of fatigue, and the impact on QoL. Continuous monitoring of pregnant women with COVID-19 is needed to develop comprehensive management strategies.
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