Search

Gastric carcinomas are among the most common cancers in Germany, with approximately 18,000 new cases per year.About 10 years ago, based on results of the Trastuzumab for gastric cancer (ToGA) trial, the addition of the monoclonal antibody trastuzumab to a platinum-fluoropyrimidine chemotherapy backbone became the standard-of-care 1st-line therapy for human epidermal growth factor receptor 2 (HER2)-positive gastric cancers. Only patients with primary HER2 gene amplification benefit from this therapy. Thus, accurate HER2 gene amplification detection is predictive and critical for therapy selection. As a gold standard the HER2 status is currently determined in tumor tissue specimens using immune histochemistry and fluorescent in situ hybridisation. However, HER2 amplification is detectable in only about 20 % of gastric carcinomas. The recent approval of an antibody-drug conjugate Trastuzumab deruxtecan (T-DXd) and the establishment of a new subgroup of HER2-low tumors due to the bystander effect associated with T-DXd increases the relevance of precise HER2 diagnostics.Aim of this analysis was to determine the HER2 amplification status from circulating DNA fragments in blood using a HER2 Copy Number Variation assay to establish a minimal invasive approach.For the present study, a digital droplet PCR-based method was validated relative to established tissue-based methods. Furthermore and most importantly, the changes of HER2 status during therapy were investigated in seven patients indicating that the changes of HER2 status and number of HER2 copies detected in blood can reflect on therapy efficiency and uncover treatment resistance.

Secondary central nervous system lymphoma (SCNSL) is a rare and difficult to treat type of Non-Hodgkin lymphoma characterized by systemic and central nervous system (CNS) disease manifestations. In this study, 124 patients with SCNSL intensively treated and with clinical long-term follow-up were included. Initial histopathology, as divided in low-grade, other aggressive, and diffuse large B-cell lymphoma (DLBCL), was of prognostic significance. Overall response to induction treatment was a prognostic factor with early responding DLBCL-SCNSL in comparison to those non-responding experiencing a significantly better progression-free survival (PFS) and overall survival (OS). However, the type of induction regime was not prognostic for survival. Following consolidating high-dose chemotherapy and autologous stem cell transplantation (HDT-ASCT), DLBCL-SCNSL patients had better median PFS and OS. The important role of HDT-ASCT was further highlighted by favorable responses and survival of patients not responding to induction therapy and by excellent results in patients with de novo DLBCL-SCNSL (65% long-term survival). SCNSL identified as a progression of disease within 6 months of initial systemic lymphoma presentation represented a previously not appreciated subgroup with particularly dismal outcome. This temporal stratification model of SCNSL diagnosis revealed CNS progression of disease within 6 months as a promising candidate prognosticator for future studies.

In the context of colorectal cancer (CRC), circulating tumor DNA (ctDNA) is frequently used to monitor the minimal residual disease (MRD). ctDNA has become an excellent biomarker to predict which patients with CRC are likely to relapse due to the persistence of micrometastases. MRD diagnosis via analysis of ctDNA may allow much earlier detection of relapse compared with conventional diagnosis during follow-up. It should lead to an increased rate of curative-intended complete resection of an asymptomatic relapse. Besides, ctDNA can provide crucial information on whether and how intensively adjuvant or additive therapy should be administered. In the present case, analysis of ctDNA gave us a crucial hint to the use of more intensive diagnostics (MRI and Positron emission tomography–computed tomography PET-CT) which led to earlier detection of CRC relapse. Metastasis detected early are more likely to be completely resectable with curative intent.

Chimeric antigen receptor (CAR)-T cells are increasingly used for the treatment of hematologic malignancies. Treatment success relies highly upon sufficient expansion of CAR-T effector cells. Accordingly, longitudinal quantification of CAR-T cells during therapy is clinically important. Techniques to quantify CAR-T cells in patient blood samples are based on flow cytometry and PCR. However, cellular kinetics of CAR-T cells are very complex and under current investigation. In this study, feasibility of CAR-T cell quantification by cell-free DNA (cfDNA) was analyzed. cfDNA isolated from 74 blood samples of 12 patients during lymphoma treatment with the anti-CD19 CAR-T cell product axicabtagene ciloleucel (axi-cel) were analyzed. Concentrations of cfDNA specific for the CAR-T gene construct (cfCAR-DNA) and a reference gene were quantified by a newly designed digital-droplet PCR (ddPCR) assay. Detection and quantification of cfCAR-DNA was feasible and reliable for all patients included. Relative quantification of cfCAR-DNA compared to a reference gene, suitable for genomic DNA analysis, was heterogeneous in treatment responders and non-responders. In contrast, parallel analyses of cfCAR-DNA and reference cfDNA in a patient-specific approach gave insight into active lymphoma killing and treatment responses. In summary, plasma cfDNA determination in lymphoma patients is a promising tool for future clinical decision making.

In acute myeloid leukemia (AML), treatment decisions are currently made according to the risk classification of the European LeukemiaNet (ELN), which is based on genetic alterations. Recently, optical genome mapping (OGM) as a novel method proved to yield a genome-wide and detailed cytogenetic characterization at the time of diagnosis. A young female patient suffered from a rather unexpected aggressive disease course under FLT3 targeted therapy in combination with induction chemotherapy. By applying a “next-generation diagnostic workup“ strategy with OGM and whole-exome sequencing (WES), a DDX3X: MLLT10 gene fusion could be detected, otherwise missed by routine diagnostics. Furthermore, several aspects of lineage ambiguity not shown by standard diagnostics were unraveled such as deletions of SUZ12 and ARPP21, as well as T-cell receptor recombination. In summary, the detection of this particular gene fusion DDX3X: MLLT10 in a female AML patient and the findings of lineage ambiguity are potential explanations for the aggressive course of disease. Our study demonstrates that OGM can yield novel clinically significant results, including additional information helpful in disease monitoring and disease biology.

In patients with compromised immune function, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and coronavirus disease 2019 (CoVID-19) impose particular challenges. Especially in hematological malignancies, including lymphoma, the demands by this novel virus disease are further enhanced during sophisticated treatments, such as chimeric antigen receptor (CAR) T-cell therapy. Here, we present the first case of a patient with refractory diffuse-large B-cell lymphoma, who underwent CAR T-cell treatment in the context of SARS-CoV-2. Irrespective of prolonged and active SARS-CoV-2 infection, T cells were successfully isolated by apheresis and processed to anti-CD19 CAR T cells (axicabtagene-ciloleucel). In light of the aggressive lymphoma course, lymphodepleting chemotherapy and CAR-T cells were administered in early recovery after oxygen-dependent CoVID-19 pneumonia. Except for moderate cytokine release, this cellular immunotherapy was well tolerated. Notably, there is no deterioration of the SARS-CoV-2 infection; however, complete lymphoma response and full clinical recovery were observed. In conclusion, CAR T-cell treatment in aggressive lymphoma in the setting of SARS-CoV-2 infection is feasible and may offer significant therapeutic activity in refractory disease.

Therapeutic strategies for patients with locally advanced rectal cancer (LARC) who are achieving a pathological complete response (pCR) after neoadjuvant radio-chemotherapy (neoCRT) are being increasingly investigated. Recent trials challenge the current standard therapy of total mesorectal excision (TME). For some patients, the treatment strategy of “watch-and-wait” seems a preferable procedure. The key factor in determining individual treatment strategies following neoCRT is the precise evaluation of the tumor response. Contrast-enhanced computer tomography (ceCT) has proven its ability to discriminate benign and malign lesions in multiple cancers. In this study, we retrospectively analyzed the ceCT based density of LARC in 30 patients, undergoing neoCRT followed by TME. We compared the tumors´ pre- and post-neoCRT density and correlated the results to the amount of residual vital tumor cells in the resected tissue. Overall, the density decreased after neoCRT, with the highest decrease in patients achieving pCR. Densitometry demonstrated a specificity of 88% and sensitivity of 68% in predicting pCR. Thus, we claim that ceCT based densitometry is a useful tool in identifying patients with LARC who may benefit from a “watch-and-wait” strategy and suggest further prospective studies.

Hemophagocytic lymphohistiocytosis (HLH) is a disorder of uncontrolled immune activation with distinct clinical features including fever, cytopenia, splenomegaly, and sepsis-like symptoms. In a young adolescent patient a novel germline GATA2 variant (NM_032638.5 (GATA2): c.177C>G, p.Tyr59Ter) was discovered and had resulted in non-tuberculous mycobacterial (NTM) infection and aggressive HLH. Strikingly, impaired degranulation of cytotoxic T-lymphocytes (CTL) and natural killer (NK)-cells was detected in CD107a-analyses. The affected patient was treated with HLA-matched unrelated alloHSCT, and subsequently all hematologic and infectious abnormalities including HLH and NTM resolved. This case supports early alloHSCT in GATA2 deficiencies as curative approach regardless of active NTM infection. Future studies on GATA2 c.177C>G, p.Tyr59*Ter might unravel its potential role in cytotoxic effector cell function and its contribution to HLH pathogenesis.

In relapsed and refractory multiple myeloma (MM), adoptive cell therapies (ACT) including CAR-T-cells are under clinical investigation. However, relapse due to T-cell exhaustion or limited persistence is an obstacle. Before ACT are considered in MM, high-dose (HD) melphalan followed by autologous stem-cell transplantation (autoSCT) has been administered in most clinical situations. Yet, the impact of HD chemotherapy on T-cells in MM with respect to ACT is unclear. In this study, T-lymphocytes’ phenotypes, expansion properties, lentiviral transduction efficacy, and gene expression were examined with special respect to patients following HD melphalan. Significant impairment of T-cells’ expansion and transduction rates could be demonstrated. Expansion was diminished due to inherent disadvantages of the predominant T-cell phenotype but restored over time. The quantitative fraction of CD27−/CD28− T-cells before expansion was predictive of T-cell yield. Following autoSCT, the transduction efficacy was reduced by disturbed lentiviral genome integration. Moreover, an unfavorable T-cell phenotype after expansion was demonstrated. In initial analyses of CD107a degranulation impaired T-cell cytotoxicity was detected in one patient following melphalan and autoSCT. The findings of our study have potential implications regarding the time point of leukapheresis for CAR-T-cell manufacturing. Our results point to a preferred interval of more than 3 months until patients should undergo cell separation for CAR-T therapy in the specific situation post-HD melphalan/autoSCT. Monitoring of CD27−/CD28− T-cells, has the potential to influence clinical decision making before apheresis in MM.

Treatment options for patients with metastatic colorectal cancer (mCRC) are limited. This particularly affects the largest group of patients with RAS mutations, who are considered ineligible for therapy with antiEGFR antibodies. In this liquid biopsy-based study, we performed the first in-depth analysis of the RAS mutational status in initially RAS-mutated patients during first-line therapy. RAS status of twelve patients with initially RAS-mutated mCRC was monitored longitudinally in 69 liquid biopsy samples. We focused on patients with stable disease (SD) or partial remission (PR) during first-line therapy (11 patients). Detection of fragmented RAS-mutated circulating cell-free tumor DNA (ctDNA) in plasma was performed by digital-droplet PCR (ddPCR) and BEAMing. Patients' total tumor masses were determined by measuring the tumor volumes using CT scan data. All patients with PR or SD at first follow-up showed a significant decrease of RAS mutational load. In ten patients (91%), the ctDNA-based RAS mutational status converted to wild-type in ddPCR and BEAMing. Remarkably, conversions were observed early after the first cycle of chemotherapy. Plasma concentration of ctDNA was controlled by determination of methylated WIF1-promotor ctDNA burden as a second tumor marker for mCRC. Persistent presence of methylated WIF1-promotor fragments confirmed the ongoing release of ctDNA during treatment. In patients with initially RAS-mutated mCRC, RAS mutations rapidly disappeared during first-line therapy in liquid biopsy, independent of type and intensity of chemotherapy and irrespective of anti-VEGF treatments. Following our results demonstrating conversion of RAS-mutational status, potential effectiveness of anti-EGFR antibodies in selected patients becomes an attractive hypothesis for future studies.

CD19-directed CAR-T-cells (CD19-CAR) have demonstrated remarkable clinical results in patients suffering from refractory or relapsed lymphoma and acute lymphoblastic leukemia. In order to further optimize follow-up, to explain treatment failure, and to control adverse events biomarkers for monitoring of response are urgently needed. Peak expansion and persistence are correlated with response rates and severity of side effects. However, no standardized method or commercially assay for CD19-CAR measurement is established yet. In this study, two primer-probe assays for digital-droplet PCR (ddPCR) were designed and subsequently explored on 54 samples collected from seven patients after CD19-CAR treatment with axi-cel over time. Detection and quantification of CAR-T-cells were feasible and reliable for all patients included. Peak expansion measured with our assay significantly correlated with the grade of neurologic adverse events but not with cytokine release syndrome. All patients with loss of CAR-signal eventually had disease progression. In summary, our novel assay allows monitoring of CAR-T-cells in vivo and may add to safety and efficacy of CAR-T treatment.

Post-transplant lymphoproliferative disease (PTLD) represents a serious complication following allogeneic hematopoietic stem cell transplantation (alloHSCT). Previously, survival rates of PTLD have improved due to the introduction of rituximab. However, reports on curative management of refractory PTLD are scarce. Today, there is no consensus how to treat rituximab-refractory PTLD, especially in highly aggressive disease. Here, we describe successful management of refractory EBV-associated PTLD, specifically DLBCL, with combined brentuximab vedotin and third-party EBV-specific T-cells in a multidisciplinary treatment approach.

Over the last decade, precise exoplanet transmission spectroscopy has revealed the atmospheres of dozens of exoplanets, driven largely by observatories like the Hubble Space Telescope. One major discovery has been the ubiquity of atmospheric aerosols, often blocking access to exoplanet chemical inventories. Tentative trends have been identified, showing that the clarity of planetary atmospheres may depend on equilibrium temperature. Previous work has often grouped dissimilar planets together in order to increase the statistical power of any trends, but it remains unclear from observed transmission spectra whether these planets exhibit the same atmospheric physics and chemistry. We present a reanalysis of a smaller, more physically similar sample of 15 exo-Neptune transmission spectra across a wide range of temperatures (200–1000 K). Using condensation cloud and hydrocarbon haze models, we find that the exo-Neptune population is best described by low cloud sedimentation efficiency ( f _sed ∼ 0.1) and high metallicity (100 × solar). There is an intrinsic scatter of ∼0.5 scale height, perhaps evidence of stochasticity in these planets’ formation processes. Observers should expect significant attenuation in transmission spectra of Neptune-size exoplanets, up to 6 scale heights for equilibrium temperatures between 500 and 800 K. With JWST's greater wavelength sensitivity, colder (

Carbon monoxide (CO) is predicted to be the dominant carbon-bearing molecule in giant planet atmospheres and, along with water, is important for discerning the oxygen and therefore carbon-to-oxygen ratio of these planets. The fundamental absorption mode of CO has a broad, double-branched structure composed of many individual absorption lines from 4.3 to 5.1 μ m, which can now be spectroscopically measured with JWST. Here we present a technique for detecting the rotational sub-band structure of CO at medium resolution with the NIRSpec G395H instrument. We use a single transit observation of the hot Jupiter WASP-39b from the JWST Transiting Exoplanet Community Early Release Science (JTEC ERS) program at the native resolution of the instrument ( R ∼ 2700) to resolve the CO absorption structure. We robustly detect absorption by CO, with an increase in transit depth of 264 ± 68 ppm, in agreement with the predicted CO contribution from the best-fit model at low resolution. This detection confirms our theoretical expectations that CO is the dominant carbon-bearing molecule in WASP-39b’s atmosphere and further supports the conclusions of low C/O and supersolar metallicities presented in the JTEC ERS papers for WASP-39b.

We present the first exoplanet phase-curve measurement made with the JWST NIRSpec instrument, highlighting the exceptional stability of this newly commissioned observatory for exoplanet climate studies. The target, WASP-121b, is an ultrahot Jupiter with an orbital period of 30.6 hr. We analyze two broadband light curves generated for the NRS1 and NRS2 detectors, covering wavelength ranges of 2.70–3.72 μ m and 3.82–5.15 μ m, respectively. Both light curves exhibit minimal systematics, with approximately linear drifts in the baseline flux level of 30 ppm hr ^−1 (NRS1) and 10 ppm hr ^−1 (NRS2). Assuming a simple brightness map for the planet described by a low-order spherical harmonic dipole, our light-curve fits suggest that the phase curve peaks coincide with orbital phases 3.°36 ± 0.°11 (NRS1) and 2.°66 ± 0.°12 (NRS2) prior to mideclipse. This is consistent with the strongest dayside emission emanating from eastward of the substellar point. We measure planet-to-star emission ratios of 3924 ± 7 ppm (NRS1) and 4924 ± 9 ppm (NRS2) for the dayside hemisphere and 136 ± 8 ppm (NRS1) and 630 ± 10 ppm (NRS2) for the nightside hemisphere. The latter nightside emission ratios translate to planetary brightness temperatures of 926 ± 12 K (NRS1) and 1122 ± 10 K (NRS2), which are low enough for a wide range of refractory condensates to form, including enstatite and forsterite. A nightside cloud deck may be blocking emission from deeper, hotter layers of the atmosphere, potentially helping to explain why cloud-free 3D general circulation model simulations systematically overpredict the nightside emission for WASP-121b.

Recent work on the characterization of small exoplanets has allowed us to accumulate growing evidence that sub-Neptunes with radii greater than ∼2.5 R _⊕ often host H _2 /He-dominated atmospheres both from measurements of their low bulk densities and from direct detections of their low mean molecular mass atmospheres. However, the smaller sub-Neptunes in the 1.5–2.2 R _⊕ size regime are much less understood and often have bulk densities that can be explained either by the H _2 /He-rich scenario or by a volatile-dominated composition known as the “water world” scenario. Here we report the detection of water vapor in the transmission spectrum of the 1.96 ± 0.08 R _⊕ sub-Neptune GJ 9827 d obtained with the Hubble Space Telescope (HST). We observe 11 HST Wide Field Camera 3 transits of GJ 9827 d and find an absorption feature at 1.4 μ m in its transit spectrum, which is best explained (at 3.39 σ ) by the presence of water in GJ 9827 d’s atmosphere. We further show that this feature cannot be caused by unocculted starspots during the transits by combining an analysis of the K2 photometry and transit light source effect retrievals. We reveal that the water absorption feature can be similarly well explained by a small amount of water vapor in a cloudy H _2 /He atmosphere or a water vapor envelope on GJ 9827 d. Given that recent studies have inferred an important mass-loss rate (>0.5 M _⊕ Gyr ^−1 ) for GJ 9827 d, making it unlikely to retain a H-dominated envelope, our findings highlight GJ 9827 d as a promising water world candidate that could host a volatile-dominated atmosphere. This water detection also makes GJ 9827 d the smallest exoplanet with an atmospheric molecular detection to date.

TOI-270 d is a temperate sub-Neptune discovered by the Transiting Exoplanet Survey Satellite (TESS) around a bright ( J = 9.1 mag) M3V host star. With an approximate radius of 2 R _⊕ and equilibrium temperature of 350 K, TOI-270 d is one of the most promising small exoplanets for atmospheric characterization using transit spectroscopy. Here we present a primary transit observation of TOI-270 d made with the Hubble Space Telescope Wide Field Camera 3 (WFC3) spectrograph across the 1.126–1.644 μ m wavelength range, and a 95% credible upper limit of 8.2 × 10 ^−14 erg s ^−1 cm ^−2 Å ^−1 arcsec ^−2 for the stellar Ly α emission obtained using the Space Telescope Imaging Spectrograph. The transmission spectrum derived from the TESS and WFC3 data provides evidence for molecular absorption by a hydrogen-rich atmosphere at 4 σ significance relative to a featureless spectrum. The strongest evidence for any individual absorber is obtained for H _2 O, which is favored at 3 σ significance. When retrieving on the WFC3 data alone and allowing for the possibility of a heterogeneous stellar brightness profile, the detection significance of H _2 O is reduced to 2.8 σ . Further observations are therefore required to robustly determine the atmospheric composition of TOI-270 d and assess the impact of stellar heterogeneity. If confirmed, our findings would make TOI-270 d one of the smallest and coolest exoplanets to date with detected atmospheric spectral features.

One of the most well-studied exoplanets to date, HD 189733 b, stands out as an archetypal hot Jupiter with many observations and theoretical models aimed at characterizing its atmosphere, interior, host star, and environment. We report here on the results of an extensive campaign to observe atmospheric escape signatures in HD 189733 b using the Hubble Space Telescope and its unique ultraviolet capabilities. We have found a tentative, but repeatable in-transit absorption of singlyionized carbon (C ii , 5.2% ± 1.4%) in the epoch of June–July/2017, as well as a neutral hydrogen (H i ) absorption consistent with previous observations. We model the hydrodynamic outflow of HD 189733 b using an isothermal Parker wind formulation to interpret the observations of escaping C and O nuclei at the altitudes probed by our observations. Our forward models indicate that the outflow of HD 189733 b is mostly neutral within an altitude of ∼2 R _p and singly ionized beyond that point. The measured in-transit absorption of C ii at 1335.7 Å is consistent with an escape rate of ∼1.1 × 10 ^11 g s ^−1 , assuming solar C abundance and an outflow temperature of 12,100 K. Although we find marginal neutral oxygen (O i ) in-transit absorption, our models predict an in-transit depth that is only comparable to the size of measurement uncertainties. A comparison between the observed Ly α transit depths and hydrodynamics models suggests that the exosphere of this planet interacts with a stellar wind at least 1 order of magnitude stronger than solar.

The search for signs of life through the detection of exoplanet atmosphere biosignature gases is gaining momentum. Yet, only a handful of rocky exoplanet atmospheres are suitable for observation with planned next-generation telescopes. To broaden prospects, we describe the possibilities for an aerial, liquid water cloud-based biosphere in the atmospheres of sub Neptune-sized temperate exoplanets, those receiving Earth-like irradiation from their host stars. One such planet is known (K2-18b) and other candidates are being followed up. Sub Neptunes are common and easier to study observationally than rocky exoplanets because of their larger sizes, lower densities, and extended atmospheres or envelopes. Yet, sub Neptunes lack any solid surface as we know it, so it is worthwhile considering whether their atmospheres can support an aerial biosphere. We review, synthesize, and build upon existing research. Passive microbial-like life particles must persist aloft in a region with liquid water clouds for long enough to metabolize, reproduce, and spread before downward transport to lower altitudes that may be too hot for life of any kind to survive. Dynamical studies are needed to flesh out quantitative details of life particle residence times. A sub Neptune would need to be a part of a planetary system with an unstable asteroid belt in order for meteoritic material to provide nutrients, though life would also need to efficiently reuse and recycle metals. The origin of life may be the most severe limiting challenge. Regardless of the uncertainties, we can keep an open mind to the search for biosignature gases as a part of general observational studies of sub Neptune exoplanets.