Effective bridging therapy can improve CD19 CAR-T outcomes while maintaining safety in patients with large B-cell lymphoma

The impact of bridging therapy (BT) on CD19-directed chimeric antigen receptor T-cell (CD19CAR-T) outcomes in large B-cell lymphoma (LBCL) is poorly characterised. Current practice is guided by physician preference rather than established evidence. Identification of effective BT modalities and factors predictive of response could improve CAR-T intention to treat and clinical outcomes. We assessed BT modality and response in 375 adult LBCL patients in relation to outcomes following axicabtagene ciloleucel (Axi-cel) or tisagenlecleucel (Tisa-cel). The majority of patients received BT with chemotherapy (57%) or radiotherapy (17%). We observed that BT was safe for patients, with minimal morbidity/mortality. We showed that complete or partial response to BT conferred a 42% reduction in disease progression and death following CD19CAR-T therapy. Multivariate analysis identified several factors associated with likelihood of response to BT, including response to last line therapy, the absence of bulky disease, and the use of Polatuzumab-containing chemotherapy regimens. Our data suggested that complete/partial response to BT may be more important for Tisa-cel than Axi-cel, as all Tisa-cel patients with less than partial response to BT experienced frank relapse within 12 months of CD19CAR-T infusion. In summary, BT in LBCL should be carefully planned towards optimal response and disease debulking, to improve CD19CAR-T patient outcomes. Polatuzumab-containing regimens should be strongly considered for all suitable patients, and failure to achieve complete/partial response to BT pre-Tisa-cel may prompt consideration of further lines of BT where possible

Persistent SARS-CoV-2 infection in patients with secondary antibody deficiency: successful clearance following combination casirivimab and imdevimab (REGN-COV2) monoclonal antibody therapy

Background: There is growing evidence that antibody responses play a role in the resolution of SARS-CoV-2 infection. Patients with primary or secondary antibody deficiency are at increased risk of persistent infection. This challenging clinical scenario is associated with adverse patient outcome and potentially creates an ecological niche for the evolution of novel SARS-CoV-2 variants with immune evasion capacity. Case reports and/or series have implied a therapeutic role for convalescent plasma (CP) to secure virological clearance, although concerns have been raised about the effectiveness of CP and its potential to drive viral evolution, and it has largely been withdrawn from clinical use in the UK. Case presentation: We report two cases in which persistent SARS-CoV-2 infection was cleared following administration of the monoclonal antibody combination casirivimab and imdevimab (REGN-COV2, Ronapreve). A 55-year-old male with follicular lymphoma, treated with B cell depleting therapy, developed SARS-CoV-2 infection in September 2020 which then persisted for over 200 days. He was hospitalised on four occasions with COVID-19 and suffered debilitating fatigue and malaise throughout. There was no clinical response to antiviral therapy with remdesivir or CP, and SARS-CoV-2 was consistently detected in nasopharyngeal swabs. Intrahost evolution of several spike variants of uncertain significance was identified by viral sequence analysis. Delivery of REGN-COV2, in combination with remdesivir, was associated with clinical improvement and viral clearance within 6 days, which was sustained for over 150 days despite immunotherapy for relapsed follicular lymphoma. The second case, a 68-year-old female with chronic lymphocytic leukaemia on ibrutinib, also developed persistent SARS-CoV-2 infection. Despite a lack of response to remdesivir, infection promptly cleared following REGN-COV2 in combination with remdesivir, accompanied by resolution of inflammation and full clinical recovery that has been maintained for over 290 days. Conclusions: These cases highlight the potential benefit of REGN-COV2 as therapy for persistent SARS-CoV-2 infection in antibody deficient individuals, including after failure of CP treatment. Formal clinical studies are warranted to assess the effectiveness of REGN-COV2 in antibody-deficient patients, especially in light of the emergence of variants of concern, such as Omicron, that appear to evade REGN-COV2 neutralisation

Single-cell Hi-C reveals cell-to-cell variability in chromosome structure.

Large-scale chromosome structure and spatial nuclear arrangement have been linked to control of gene expression and DNA replication and repair. Genomic techniques based on chromosome conformation capture (3C) assess contacts for millions of loci simultaneously, but do so by averaging chromosome conformations from millions of nuclei. Here we introduce single-cell Hi-C, combined with genome-wide statistical analysis and structural modelling of single-copy X chromosomes, to show that individual chromosomes maintain domain organization at the megabase scale, but show variable cell-to-cell chromosome structures at larger scales. Despite this structural stochasticity, localization of active gene domains to boundaries of chromosome territories is a hallmark of chromosomal conformation. Single-cell Hi-C data bridge current gaps between genomics and microscopy studies of chromosomes, demonstrating how modular organization underlies dynamic chromosome structure, and how this structure is probabilistically linked with genome activity patterns

The Vehicle, Spring 1974

Table of Contents PhotoJim Painterpage 1 Six Poems of the LandRay Schmuddepage 5 At Last to Find FreedomJann Briesacherpage 7 The Last IrisMarjorie Thoelepage 9 (Untitled)Melinda E. Recordpage 10 MenJan Schroederpage 10 ImpressionsJudy Bardpage 11 ScaredAnita Surpage 11 Loved and LostJan Schroederpage 12 Dripped-Over WaxAnita Surpage 13 The Crowded RoomWilliam E. Uteschpage 14 A River in IllinoisJames Jonespage 14 Sneeze SeasonDarlene A. Moorepage 14 ChangesMark Chianakaspage 15 PhotoJim Painterpage 16 Wedding VowsJann Briesacherpage 17 PhotoJim Painterpage 18 PhotoJim Painterpage 19 PhotoJim Painterpage 20 PhotoJim Painterpage 21 PhotoJim Painterpage 22 PhotoLarry Smyserpage 23 From Outside ColoradoRay Schmuddepage 24 Dairy QueenGayle Gleichmanpage 26 With Sunstreaks in our HairNancy Broom Brownpage 33 PhotoJim Painterpage 34 Water\u27s EdgeMarjorie Thoelepage 35 My 665th Illusion of SanityGordon Glessnerpage 36 Is it my turn to do the laundry again??? Jann Briesacherpage 38 TV Teachingbobbdoddpage 39 GuidanceWendy Diane Wielandpage 40 PhotoJim Painterpage 41 RaindropsJane Ann Beerspage 42 WaitingJan Schroederpage 42 To JonJudy Bardpage 43 One Autumn Day in 1971E. Christmanpage 43 More Surely Than Picture AlbumsMarjorie Thoelepage 44 WingspanningNancy Broom Brownpage 45 ReligionMelinda E. Recordpage 45 Rosalie StevensonMark Holleypage 46 PhotoJim Painterpage 47 WhiteShirley A. Rardinpage 48 The Beginning of a Perfect DayShirley A. Rardinpage 49 PhotoMichael Chenpage 50 Rosethorn Wall of June 17bobbdoddpage 51 ManJan Schroederpage 51 HaikuJudy Bardpage 51 You know it leaves me emptyJames Osbornepage 52 For JesseJames Osbornepage 52 EndingsMark Chianakaspage 53 ConfusionGary L. Owenspage 53 PhotoMichael Chenpage 54 PoemsJann Briesacherpage 54 Journey of just oneNancy Broom Brownpage 55 Blackbirds in IllinoisJames Jonespage 56 PoemsJann Briesacherpage 56 PhotoMichael Chenpage 57 I am a poemDarlene A. Moorepage 57 A Glimpse of ParadiseJann Briesacherpage 57 PhotoJim Painterpage 58 PoemSheila Marie Foorpage 59 In my windowBarbara S. Meyerpage 59 Section 4., Draft 3bobbdoddpage 60 PhotoJim Painterpage 61 PoemJann Briesacherpage 61 PhotoGary Deanpage 62 I amWilliam E. Uteschpage 62 To a tank-car in IllinoisJames Jonespage 63 PoemJane Ann Beerspage 63 PoemsJann Briesacherpage 63 Editor\u27s Pagepage 64https://thekeep.eiu.edu/vehicle/1031/thumbnail.jp

Dual targeting of CD19 and CD22 with Bicistronic CAR-T cells in Patients with Relapsed/Refractory Large B Cell Lymphoma

Relapse following CD19-directed chimeric antigen receptor T-cells (CAR-T) for relapsed/refractory large B-cell lymphoma (r/r LBCL) is commonly ascribed to antigen loss or CAR-T exhaustion. Multi-antigen targeting and PD-1 blockade are rational approaches to prevent relapse. Here, we test CD19/22 dual-targeting CAR-T (AUTO3) plus pembrolizumab in r/r LBCL as inpatient or outpatient therapy (NCT03289455, https://clinicaltrials.gov/ct2/show/NCT03289455). Endpoints include toxicity (primary) and response rates (secondary). AUTO3 was manufactured for 62 patients using autologous leukapheresis, modified with a bicistronic transgene. 52 patients received AUTO3 (7/52,50x106; 45/52,150-450x106) and 48/52 received pembrolizumab. Median age was 59 years (range,27-83) and 46/52 had stage III/IV disease. Median follow-up was 21.6 months (range,15.1-51.3) at last data cut (Feb 28, 2022). AUTO3 was safe: grade 1-2 and grade 3 CRS affected 18/52 (34.6%) and 1/52 (1.9%) patients, neurotoxicity arose in 4 patients (2/4, grade 3-4), HLH affected 2 patients, and no Pembrolizumab-associated autoimmune sequalae were observed. On this basis, outpatient administration was tested in 20 patients, saving a median of 14 hospital days/patient. AUTO3 was effective: overall response rates were 66% (48.9%, CR; 17%, PR). For patients with CR, median DOR was not reached, with 54.4% (CI: 32.8, 71.7) projected to remain progression-free beyond 12 months after onset of remission. DOR for all responding patients was 8.3 months (95% CI: 3.0, NE) with 42.6% projected to remain progression-free beyond 12 months after onset of remission. Overall, AUTO3 +/- pembrolizumab for r/r LBCL was safe, lending itself to outpatient administration, and delivered durable remissions in 54.4% of complete responders, associated with robust CAR-T expansion. Neither dual-targeting CAR-T nor pembrolizumab prevented relapse in a significant proportion of patients, and future developments include next-generation-AUTO3, engineered for superior expansion/persistence in vivo, and selection of CAR binders active at low antigen densities

The Vehicle, Spring 1974

Table of Contents PhotoJim Painterpage 1 Six Poems of the LandRay Schmuddepage 5 At Last to Find FreedomJann Briesacherpage 7 The Last IrisMarjorie Thoelepage 9 (Untitled)Melinda E. Recordpage 10 MenJan Schroederpage 10 ImpressionsJudy Bardpage 11 ScaredAnita Surpage 11 Loved and LostJan Schroederpage 12 Dripped-Over WaxAnita Surpage 13 The Crowded RoomWilliam E. Uteschpage 14 A River in IllinoisJames Jonespage 14 Sneeze SeasonDarlene A. Moorepage 14 ChangesMark Chianakaspage 15 PhotoJim Painterpage 16 Wedding VowsJann Briesacherpage 17 PhotoJim Painterpage 18 PhotoJim Painterpage 19 PhotoJim Painterpage 20 PhotoJim Painterpage 21 PhotoJim Painterpage 22 PhotoLarry Smyserpage 23 From Outside ColoradoRay Schmuddepage 24 Dairy QueenGayle Gleichmanpage 26 With Sunstreaks in our HairNancy Broom Brownpage 33 PhotoJim Painterpage 34 Water\u27s EdgeMarjorie Thoelepage 35 My 665th Illusion of SanityGordon Glessnerpage 36 Is it my turn to do the laundry again??? Jann Briesacherpage 38 TV Teachingbobbdoddpage 39 GuidanceWendy Diane Wielandpage 40 PhotoJim Painterpage 41 RaindropsJane Ann Beerspage 42 WaitingJan Schroederpage 42 To JonJudy Bardpage 43 One Autumn Day in 1971E. Christmanpage 43 More Surely Than Picture AlbumsMarjorie Thoelepage 44 WingspanningNancy Broom Brownpage 45 ReligionMelinda E. Recordpage 45 Rosalie StevensonMark Holleypage 46 PhotoJim Painterpage 47 WhiteShirley A. Rardinpage 48 The Beginning of a Perfect DayShirley A. Rardinpage 49 PhotoMichael Chenpage 50 Rosethorn Wall of June 17bobbdoddpage 51 ManJan Schroederpage 51 HaikuJudy Bardpage 51 You know it leaves me emptyJames Osbornepage 52 For JesseJames Osbornepage 52 EndingsMark Chianakaspage 53 ConfusionGary L. Owenspage 53 PhotoMichael Chenpage 54 PoemsJann Briesacherpage 54 Journey of just oneNancy Broom Brownpage 55 Blackbirds in IllinoisJames Jonespage 56 PoemsJann Briesacherpage 56 PhotoMichael Chenpage 57 I am a poemDarlene A. Moorepage 57 A Glimpse of ParadiseJann Briesacherpage 57 PhotoJim Painterpage 58 PoemSheila Marie Foorpage 59 In my windowBarbara S. Meyerpage 59 Section 4., Draft 3bobbdoddpage 60 PhotoJim Painterpage 61 PoemJann Briesacherpage 61 PhotoGary Deanpage 62 I amWilliam E. Uteschpage 62 To a tank-car in IllinoisJames Jonespage 63 PoemJane Ann Beerspage 63 PoemsJann Briesacherpage 63 Editor\u27s Pagepage 64https://thekeep.eiu.edu/vehicle/1031/thumbnail.jp

Activation of Estrogen-Responsive Genes Does Not Require Their Nuclear Co-Localization

The spatial organization of the genome in the nucleus plays a role in the regulation of gene expression. Whether co-regulated genes are subject to coordinated repositioning to a shared nuclear space is a matter of considerable interest and debate. We investigated the nuclear organization of estrogen receptor alpha (ERα) target genes in human breast epithelial and cancer cell lines, before and after transcriptional activation induced with estradiol. We find that, contrary to another report, the ERα target genes TFF1 and GREB1 are distributed in the nucleoplasm with no particular relationship to each other. The nuclear separation between these genes, as well as between the ERα target genes PGR and CTSD, was unchanged by hormone addition and transcriptional activation with no evidence for co-localization between alleles. Similarly, while the volume occupied by the chromosomes increased, the relative nuclear position of the respective chromosome territories was unaffected by hormone addition. Our results demonstrate that estradiol-induced ERα target genes are not required to co-localize in the nucleus

MATRix-RICE therapy and autologous haematopoietic stem-cell transplantation in diffuse large B-cell lymphoma with secondary CNS involvement (MARIETTA): an international, single-arm, phase 2 trial.

BACKGROUND Secondary CNS lymphoma is a rare but potentially lethal event in patients with diffuse large B-cell lymphoma. We aimed to assess the activity and safety of an intensive, CNS-directed chemoimmunotherapy consolidated by autologous haematopoietic stem-cell transplantation (HSCT) in patients with secondary CNS lymphoma. METHODS This international, single-arm, phase 2 trial was done in 24 hospitals in Italy, the UK, the Netherlands, and Switzerland. Adults (aged 18-70 years) with histologically diagnosed diffuse large B-cell lymphoma and CNS involvement at the time of primary diagnosis or at relapse and Eastern Cooperative Oncology Group Performance Status of 3 or less were enrolled and received three courses of MATRix (rituximab 375 mg/m2, intravenous infusion, day 0; methotrexate 3·5 g/m2, the first 0·5 g/m2 in 15 min followed by 3 g/m2 in a 3 h intravenous infusion, day 1; cytarabine 2 g/m2 every 12 h, in 1 h intravenous infusions, days 2 and 3; thiotepa 30 mg/m2, 30 min intravenous infusion, day 4) followed by three courses of RICE (rituximab 375 mg/m2, day 1; etoposide 100 mg/m2 per day in 500-1000 mL over a 60 min intravenous infusion, days 1, 2, and 3; ifosfamide 5 g/m2 in 1000 mL in a 24 h intravenous infusion with mesna support, day 2; carboplatin area under the curve of 5 in 500 mL in a 1 h intravenous infusion, day 2) and carmustine-thiotepa and autologous HSCT (carmustine 400 mg/m2 in 500 mL glucose 5% solution in a 1-2 h infusion, day -6; thiotepa 5 mg/kg in saline solution in a 2 h infusion every 12 h, days -5 and -4). The primary endpoint was progression-free survival at 1 year. Overall and complete response rates before autologous HSCT, duration of response, overall survival, and safety were the secondary endpoints. Analyses were in the modified intention-to-treat population. This study is registered with ClinicalTrials.gov, NCT02329080. The trial ended after accrual completion; the database lock was Dec 31, 2019. FINDINGS Between March 30, 2015, and Aug 3, 2018, 79 patients were enrolled. 75 patients were assessable. 319 (71%) of the 450 planned courses were delivered. At 1 year from enrolment the primary endpoint was met, 42 patients were progression free (progression-free survival 58%; 95% CI 55-61). 49 patients (65%; 95% CI 54-76) had an objective response after MATRix-RICE, 29 (39%) of whom had a complete response. 37 patients who responded had autologous HSCT. At the end of the programme, 46 patients (61%; 95% CI 51-71) had an objective response, with a median duration of objective response of 26 months (IQR 16-37). At a median follow-up of 29 months (IQR 20-40), 35 patients were progression-free and 33 were alive, with a 2-year overall survival of 46% (95% CI 39-53). Grade 3-4 toxicity was most commonly haematological: neutropenia in 46 (61%) of 75 patients, thrombocytopenia in 45 (60%), and anaemia in 26 (35%). 79 serious adverse events were recorded in 42 (56%) patients; four (5%) of those 79 were lethal due to sepsis caused by Gram-negative bacteria (treatment-related mortality 5%; 95% CI 0·07-9·93). INTERPRETATION MATRix-RICE plus autologous HSCT was active in this population of patients with very poor prognosis, and had an acceptable toxicity profile. FUNDING Stand Up To Cancer Campaign for Cancer Research UK, the Swiss Cancer Research foundation, and the Swiss Cancer League

Changes in chromatin structure during processing of wax-embedded tissue sections

The use of immunofluorescence (IF) and fluorescence in situ hybridisation (FISH) underpins much of our understanding of how chromatin is organised in the nucleus. However, there has only recently been an appreciation that these types of study need to move away from cells grown in culture and towards an investigation of nuclear organisation in cells in situ in their normal tissue architecture. Such analyses, however, especially of archival clinical samples, often requires use of formalin-fixed paraffin wax-embedded tissue sections which need addition steps of processing prior to IF or FISH. Here we quantify the changes in nuclear and chromatin structure that may be caused by these additional processing steps. Treatments, especially the microwaving to reverse fixation, do significantly alter nuclear architecture and chromatin texture, and these must be considered when inferring the original organisation of the nucleus from data collected from wax-embedded tissue sections