CAR-T cell therapy has revolutionized the treatment of non-Hodgkin B-lymphomas, but relapse rates remain unacceptably high, and resistance mechanisms are still unclear. A key challenge of T-cell−based therapy is the immune-suppressive tumor microenvironment, which is rich in immune-suppressive cells, including regulatory T cells (Tregs). Tregs express FOXP3, CD3 and CD25, and their role in B-cell lymphomas treated with CAR-T cells remains uncertain. Using immunohistochemistry, we analyzed 31 pre-infusion and seven post-infusion paired biopsies from patients with relapsed/refractory B-cell lymphoma who received CAR-T cell therapy. Pre- infusion FOXP3+ cell percentages were assessed and correlated with response rate, progression-free survival (PFS), and overall survival (OS). Post-infusion biopsies were examined for FOXP3 changes. We identified 3% as the optimal FOXP3+ cut-off, defining low (≤3%) and high (>3%) FOXP3 groups. Patients with high FOXP3 had a better OS (1-year OS 94% versus 61%, P = 0.006). PFS also favored high FOXP3 patients but lacked statistical significance (1-year PFS 65% versus 54%, P = 0.41). Notably, relapsed patients with high FOXP3 maintained high expression and responded better to subsequent treatments (P = 0.03). Our findings highlight the impact of tumor-infiltrating Tregs on CAR-T cell efficacy in lymphomas.
CAR-T细胞疗法已经彻底改变了非霍奇金b淋巴瘤的治疗，但复发率仍然高得令人无法接受，而且耐药机制仍不清楚。基于T细胞的治疗的一个关键挑战是免疫抑制肿瘤微环境，其中富含免疫抑制细胞，包括调节性T细胞（Tregs）。Tregs表达FOXP3、CD3和CD25，它们在CAR-T细胞治疗的b细胞淋巴瘤中的作用尚不确定。使用免疫组织化学，我们分析了31例输注前和7例输注后的成对活检，这些活检来自接受CAR-T细胞治疗的复发/难治性b细胞淋巴瘤患者。评估输注前FOXP3+细胞百分比，并将其与缓解率、无进展生存期（PFS）和总生存期（OS）相关。注射后活检检查FOXP3变化。我们确定3%为最佳FOXP3+临界值，定义了低（≤3%）和高（低于3%）FOXP3组。FOXP3高的患者有更好的OS（1年OS 94% vs 61%， P = 0.006）。PFS也有利于高FOXP3患者，但缺乏统计学意义（1年PFS 65% vs 54%， P = 0.41）。值得注意的是，FOXP3高表达的复发患者保持高表达，对后续治疗的反应更好（P = 0.03）。我们的研究结果强调了肿瘤浸润性Tregs对CAR-T细胞治疗淋巴瘤疗效的影响。
{"title":"A FOXP3+ cell-rich microenvironment associates with better overall survival after CAR-T cell therapy in relapsed/refractory B-cell lymphoma","authors":"Simone Zanella , Cristina Zucchinetti , Chiara de Philippis , Daniele Mannina , Daniela Taurino , Jacopo Mariotti , Barbara Sarina , Daoud Rahal , Carmelo Carlo-Stella , Armando Santoro , Arturo Bonometti , Stefania Bramanti","doi":"10.1016/j.jcyt.2025.10.007","DOIUrl":"10.1016/j.jcyt.2025.10.007","url":null,"abstract":"<div><div>CAR-T cell therapy has revolutionized the treatment of non-Hodgkin B-lymphomas, but relapse rates remain unacceptably high, and resistance mechanisms are still unclear. A key challenge of T-cell−based therapy is the immune-suppressive tumor microenvironment, which is rich in immune-suppressive cells, including regulatory T cells (Tregs). Tregs express FOXP3, CD3 and CD25, and their role in B-cell lymphomas treated with CAR-T cells remains uncertain. Using immunohistochemistry, we analyzed 31 pre-infusion and seven post-infusion paired biopsies from patients with relapsed/refractory B-cell lymphoma who received CAR-T cell therapy. Pre- infusion FOXP3+ cell percentages were assessed and correlated with response rate, progression-free survival (PFS), and overall survival (OS). Post-infusion biopsies were examined for FOXP3 changes. We identified 3% as the optimal FOXP3+ cut-off, defining low (≤3%) and high (>3%) FOXP3 groups. Patients with high FOXP3 had a better OS (1-year OS 94% versus 61%, <em>P</em> = 0.006). PFS also favored high FOXP3 patients but lacked statistical significance (1-year PFS 65% versus 54%, <em>P</em> = 0.41). Notably, relapsed patients with high FOXP3 maintained high expression and responded better to subsequent treatments (<em>P</em> = 0.03). Our findings highlight the impact of tumor-infiltrating Tregs on CAR-T cell efficacy in lymphomas.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"28 3","pages":"Article 101997"},"PeriodicalIF":3.2,"publicationDate":"2025-11-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145524579","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-01DOI: 10.1016/j.jcyt.2025.10.005
Jeremy Snyder
Background Aims
Cellular and gene therapy products provide hope to people experiencing debilitating diseases. Many people seek to access these products ahead of regulatory approval through clinical trials, exceptional access programs, and direct to consumer purchases. Even when regulatory approval has been given, these products can cost millions of dollars in direct and indirect expenses. This study examines how crowdfunding campaigns are being used to pay for United States Federal Drug Administration (FDA) approved cellular and gene therapy products.
Methods
The author searched the GoFundMe crowdfunding platform for 43 FDA-approved cellular and gene therapy products. The campaign title, campaign text, funding requested, funding pledged, number of donations, campaign start date, and campaign location was collected for each campaign. Total 529 campaigns were identified and 322 included for content analysis as seeking funding to pay for the direct or indirect costs of accessing FDA-approved cellular or gene therapy products.
Results
Crowdfunding campaigns for 17 FDA-approved cellular and gene therapy products were identified. Total 322 campaigns raised $8,997,107.19 (median $3675.04) from 154,230 (72.5) donations out of $334,723,452.76 ($184,250) requested. Two campaigns raised 39.2% of all funds and 10 raised 64.8% of all funds. Fundraising goals ranged from $1.11–$12,000,000. The 4th and 5th largest quintiles of fundraising requests were over $1,200,000.
Conclusions
The median fundraising goal and funding received were larger than has been found in other studies of medical crowdfunding. Differences in fundraising goals and donations suggest inequities in insurance coverage and the ability to access cellular and gene therapy products.
{"title":"Crowdfunding for cellular and gene therapy products","authors":"Jeremy Snyder","doi":"10.1016/j.jcyt.2025.10.005","DOIUrl":"10.1016/j.jcyt.2025.10.005","url":null,"abstract":"<div><h3>Background Aims</h3><div>Cellular and gene therapy products provide hope to people experiencing debilitating diseases. Many people seek to access these products ahead of regulatory approval through clinical trials, exceptional access programs, and direct to consumer purchases. Even when regulatory approval has been given, these products can cost millions of dollars in direct and indirect expenses. This study examines how crowdfunding campaigns are being used to pay for United States Federal Drug Administration (FDA) approved cellular and gene therapy products.</div></div><div><h3>Methods</h3><div>The author searched the GoFundMe crowdfunding platform for 43 FDA-approved cellular and gene therapy products. The campaign title, campaign text, funding requested, funding pledged, number of donations, campaign start date, and campaign location was collected for each campaign. Total 529 campaigns were identified and 322 included for content analysis as seeking funding to pay for the direct or indirect costs of accessing FDA-approved cellular or gene therapy products.</div></div><div><h3>Results</h3><div>Crowdfunding campaigns for 17 FDA-approved cellular and gene therapy products were identified. Total 322 campaigns raised $8,997,107.19 (median $3675.04) from 154,230 (72.5) donations out of $334,723,452.76 ($184,250) requested. Two campaigns raised 39.2% of all funds and 10 raised 64.8% of all funds. Fundraising goals ranged from $1.11–$12,000,000. The 4<sup>th</sup> and 5<sup>th</sup> largest quintiles of fundraising requests were over $1,200,000.</div></div><div><h3>Conclusions</h3><div>The median fundraising goal and funding received were larger than has been found in other studies of medical crowdfunding. Differences in fundraising goals and donations suggest inequities in insurance coverage and the ability to access cellular and gene therapy products.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"28 1","pages":"Article 101995"},"PeriodicalIF":3.2,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145580047","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-29DOI: 10.1016/j.jcyt.2025.10.008
Inês Serrenho , Sofia C. Serra , António J. Salgado , Graça Baltazar
Background aims
Neonatal hypoxic-ischemic encephalopathy (HIE) remains a leading cause of infant mortality and long-term neurologic disability. Although therapeutic hypothermia (TH) is the current standard treatment, its effectiveness is limited, with many infants either ineligible or showing incomplete recovery. As a result, alternative therapies, such as mesenchymal stem cells (MSCs) derived from induced pluripotent stem cells (iMSCs), are being explored. iMSCs have shown considerable promise in preclinical studies because of their consistent properties, which help overcome some of the challenges associated with traditional MSCs. This study evaluated the efficacy of iMSCs and their secretome in promoting recovery after neonatal hypoxic-ischemic (HI) brain injury in a well-established rodent model.
Methods
The Rice-Vannucci model was used to induce HI brain injury to the developing brain on postnatal day 10 (P10) Wistar Han rat pups. Two days post-injury, 50 000 iMSCs or their secretome were administered intranasally. Over the next 30 days, motor and cognitive functions were assessed through a series of behavioral tests. In addition, brain lesion size, neurogenesis and glial reactivity were examined by immunohistochemistry to evaluate the extent of neurorepair and inflammation.
Results
HI-lesioned animals treated with intranasal iMSCs or their secretome demonstrated improved motor capabilities and enhanced recognition memory compared with untreated lesioned animals. Both iMSCs and their secretome administration led to a reduction in brain lesion size and increased neurogenesis in the hippocampus. Moreover, glial reactivity, including astrocyte and microglia activation, was significantly reduced 30 days after injury, suggesting a more favorable neuroinflammatory environment in treated groups.
Conclusions
These findings highlight the potential of iMSCs and their secretome to enhance functional recovery and reduce brain damage after neonatal HI. The similar therapeutic outcomes achieved with the iMSC secretome suggest that the secreted factors play a central role in driving neurorepair. More studies are still necessary to gain a better understanding of the mechanisms underlying iMSCs' neuroprotective and neuroreparative effects.
{"title":"Brain on the mend: induced mesenchymal stem cell−based therapies promote functional recovery in rats with neonatal hypoxic-ischemic brain injury","authors":"Inês Serrenho , Sofia C. Serra , António J. Salgado , Graça Baltazar","doi":"10.1016/j.jcyt.2025.10.008","DOIUrl":"10.1016/j.jcyt.2025.10.008","url":null,"abstract":"<div><h3>Background aims</h3><div>Neonatal hypoxic-ischemic encephalopathy (HIE) remains a leading cause of infant mortality and long-term neurologic disability. Although therapeutic hypothermia (TH) is the current standard treatment, its effectiveness is limited, with many infants either ineligible or showing incomplete recovery. As a result, alternative therapies, such as mesenchymal stem cells (MSCs) derived from induced pluripotent stem cells (iMSCs), are being explored. iMSCs have shown considerable promise in preclinical studies because of their consistent properties, which help overcome some of the challenges associated with traditional MSCs. This study evaluated the efficacy of iMSCs and their secretome in promoting recovery after neonatal hypoxic-ischemic (HI) brain injury in a well-established rodent model.</div></div><div><h3>Methods</h3><div>The Rice-Vannucci model was used to induce HI brain injury to the developing brain on postnatal day 10 (P10) Wistar Han rat pups. Two days post-injury, 50 000 iMSCs or their secretome were administered intranasally. Over the next 30 days, motor and cognitive functions were assessed through a series of behavioral tests. In addition, brain lesion size, neurogenesis and glial reactivity were examined by immunohistochemistry to evaluate the extent of neurorepair and inflammation.</div></div><div><h3>Results</h3><div>HI-lesioned animals treated with intranasal iMSCs or their secretome demonstrated improved motor capabilities and enhanced recognition memory compared with untreated lesioned animals. Both iMSCs and their secretome administration led to a reduction in brain lesion size and increased neurogenesis in the hippocampus. Moreover, glial reactivity, including astrocyte and microglia activation, was significantly reduced 30 days after injury, suggesting a more favorable neuroinflammatory environment in treated groups.</div></div><div><h3>Conclusions</h3><div>These findings highlight the potential of iMSCs and their secretome to enhance functional recovery and reduce brain damage after neonatal HI. The similar therapeutic outcomes achieved with the iMSC secretome suggest that the secreted factors play a central role in driving neurorepair. More studies are still necessary to gain a better understanding of the mechanisms underlying iMSCs' neuroprotective and neuroreparative effects.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"28 3","pages":"Article 101998"},"PeriodicalIF":3.2,"publicationDate":"2025-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145551696","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-25DOI: 10.1016/j.jcyt.2025.10.006
Jonida Kokiçi , Helena Arellano-Ballestero , Benjamin Hammond , Anucha Preechanukul , Noshin Hussain , Kelly da Costa , Jessica Davies , Sadiyah Mukhtar , Thomas Fernandez , Sabine Kinloch , Fiona M. Burns , Patrick Kennedy , Douglas MacDonald , Mala K. Maini , Upkar S. Gill , Alan Xiaodong Zhuang , Mark W. Lowdell , Karl-Johan Malmberg , Ebba Sohlberg , Dimitra Peppa
Background
Hepatitis B virus (HBV) infection remains a significant global health challenge, leading to chronic liver disease and hepatocellular carcinoma (HCC). Natural killer (NK) cells play an important role in the clearance of HBV-infected cells, but their efficacy is often compromised during chronic infection. Adaptive NK cells, characterized by NKG2C expression and enhanced functional responses, represent a promising therapeutic avenue for enhancing anti-HBV immunity and responses to HBV-driven cancers.
Methods
We applied an established protocol, involving K562-HLA-E expressing feeder cells and cytokines (IL-2), for the expansion of adaptive NK cells from cryopreserved T- and B cell depleted peripheral blood mononuclear cells (PBMCs) derived from donors with chronic HBV infection alone or with Human Immunodeficiency Virus (HIV) co-infection. We evaluated the adaptive profile of expanded NK cells, their antibody-dependent cellular cytotoxicity (ADCC) capacity and functional responses against hepatoma cell lines in the presence or absence of HBV infection.
Results
Expanded NK cells achieved >97% purity, with the NKG2C positive population exhibiting a mean 100-fold expansion. These cells demonstrated a predominantly adaptive phenotype with high surface expression of NKG2C and cytotoxic potential (Granzyme B). They maintained high levels of CD16 surface expression and upregulated CD2, essential for ADCC. Functionally, expanded adaptive NK cells showed enhanced ADCC capacity and functional responses to K562 targets, naive, HBV integrant-expressing, and de novo infected hepatoma cell lines. TGF-β preconditioning induced tissue-resident features (CD103, CD49a) in expanded adaptive NK cells, while preserving their adaptive phenotype and functionality, enhancing their potential for liver targeted immunotherapy. Further, expanded adaptive NK cells demonstrated minimal reactivity against autologous activated T cells, suggesting limited off-target effects.
Conclusions
Our study demonstrates the first successful expansion of adaptive NK cells with robust functional responses from donors with chronic viral infection. This approach creates opportunities for NK cell-based therapies alone or in combination with monoclonal antibodies contributing to HBV functional cure strategies and the treatment of HBV-driven cancers.
{"title":"Expanded adaptive NKG2C+ NK cells exhibit potent ADCC and functional responses against HBV-infected hepatoma cell lines","authors":"Jonida Kokiçi , Helena Arellano-Ballestero , Benjamin Hammond , Anucha Preechanukul , Noshin Hussain , Kelly da Costa , Jessica Davies , Sadiyah Mukhtar , Thomas Fernandez , Sabine Kinloch , Fiona M. Burns , Patrick Kennedy , Douglas MacDonald , Mala K. Maini , Upkar S. Gill , Alan Xiaodong Zhuang , Mark W. Lowdell , Karl-Johan Malmberg , Ebba Sohlberg , Dimitra Peppa","doi":"10.1016/j.jcyt.2025.10.006","DOIUrl":"10.1016/j.jcyt.2025.10.006","url":null,"abstract":"<div><h3>Background</h3><div>Hepatitis B virus (HBV) infection remains a significant global health challenge, leading to chronic liver disease and hepatocellular carcinoma (HCC). Natural killer (NK) cells play an important role in the clearance of HBV-infected cells, but their efficacy is often compromised during chronic infection. Adaptive NK cells, characterized by NKG2C expression and enhanced functional responses, represent a promising therapeutic avenue for enhancing anti-HBV immunity and responses to HBV-driven cancers.</div></div><div><h3>Methods</h3><div>We applied an established protocol, involving K562-HLA-E expressing feeder cells and cytokines (IL-2), for the expansion of adaptive NK cells from cryopreserved T- and B cell depleted peripheral blood mononuclear cells (PBMCs) derived from donors with chronic HBV infection alone or with Human Immunodeficiency Virus (HIV) co-infection. We evaluated the adaptive profile of expanded NK cells, their antibody-dependent cellular cytotoxicity (ADCC) capacity and functional responses against hepatoma cell lines in the presence or absence of HBV infection.</div></div><div><h3>Results</h3><div>Expanded NK cells achieved >97% purity, with the NKG2C positive population exhibiting a mean 100-fold expansion. These cells demonstrated a predominantly adaptive phenotype with high surface expression of NKG2C and cytotoxic potential (Granzyme B). They maintained high levels of CD16 surface expression and upregulated CD2, essential for ADCC. Functionally, expanded adaptive NK cells showed enhanced ADCC capacity and functional responses to K562 targets, naive, HBV integrant-expressing, and <em>de novo</em> infected hepatoma cell lines. TGF-β preconditioning induced tissue-resident features (CD103, CD49a) in expanded adaptive NK cells, while preserving their adaptive phenotype and functionality, enhancing their potential for liver targeted immunotherapy. Further, expanded adaptive NK cells demonstrated minimal reactivity against autologous activated T cells, suggesting limited off-target effects.</div></div><div><h3>Conclusions</h3><div>Our study demonstrates the first successful expansion of adaptive NK cells with robust functional responses from donors with chronic viral infection. This approach creates opportunities for NK cell-based therapies alone or in combination with monoclonal antibodies contributing to HBV functional cure strategies and the treatment of HBV-driven cancers.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"28 3","pages":"Article 101996"},"PeriodicalIF":3.2,"publicationDate":"2025-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145710268","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Natural killer (NK) cell therapies hold great promise for cancer treatment; however, donor-to-donor heterogeneity in the ex vivo expansion process remains a critical bottleneck in their supply. This study aimed to identify factors influencing donor variability in a 2-week-long ex vivo NK cell expansion from peripheral blood mononuclear cells, analyzed across three donors. Single-cell transcriptomics was applied to investigate the distribution of cell types and phenotypes, as well as trajectory inference and differential gene expression. Our results identified that several factors were associated with the variability in the final NK cell fraction and expansion, and that their influence was prevalent between culture days 3 and 8. Compared to a high final NK cell expansion, a culture with a low final NK cell expansion exhibited an upregulation of some stress and inflammatory genes and an increase in one specific subcluster of NK cells already on culture day 3. It showed a low score of CD56Bright CD16– phenotype and a high score of CD56Dim CD16+ phenotype. It also had a decreased presence of cytotoxic CD8+ Tm cells. Among the observed subclusters of CD8+ Tm cells, it exhibited a higher presence of a subcluster associated with a less differentiated and less cytotoxic phenotype, as well as a lower prevalence of a subcluster associated with chemokine and cytotoxic genes. Finally, it had a major expansion of one of the CD8+ Tm cells subclusters annotated as NK-like T cell and characterized by a high CCR5 mRNA expression, while the levels of CCL3, CCL4 and CCL5 mRNA were downregulated. The present findings point toward a potential link between CCL signaling and improved NK cell expansion performance, including possible markers for further investigations, and suggest future strategies to increase the final NK cell fraction and expansion based on donor-specific markers.
{"title":"Single-cell transcriptomics reveals dynamics of natural killer cell expansion in a feeder cell-free culture of peripheral blood mononuclear cells—implications for immunotherapy","authors":"Brian Ladd , Markella Zacharouli , Per-Henrik Holmqvist , Stefanie Renken , Pontus Blomberg , Véronique Chotteau","doi":"10.1016/j.jcyt.2025.10.004","DOIUrl":"10.1016/j.jcyt.2025.10.004","url":null,"abstract":"<div><div>Natural killer (NK) cell therapies hold great promise for cancer treatment; however, donor-to-donor heterogeneity in the <em>ex vivo</em> expansion process remains a critical bottleneck in their supply. This study aimed to identify factors influencing donor variability in a 2-week-long <em>ex vivo</em> NK cell expansion from peripheral blood mononuclear cells, analyzed across three donors. Single-cell transcriptomics was applied to investigate the distribution of cell types and phenotypes, as well as trajectory inference and differential gene expression. Our results identified that several factors were associated with the variability in the final NK cell fraction and expansion, and that their influence was prevalent between culture days 3 and 8. Compared to a high final NK cell expansion, a culture with a low final NK cell expansion exhibited an upregulation of some stress and inflammatory genes and an increase in one specific subcluster of NK cells already on culture day 3. It showed a low score of CD56<sup>Bright</sup> CD16<sup>–</sup> phenotype and a high score of CD56<sup>Dim</sup> CD16<sup>+</sup> phenotype. It also had a decreased presence of cytotoxic CD8<sup>+</sup> Tm cells. Among the observed subclusters of CD8<sup>+</sup> Tm cells, it exhibited a higher presence of a subcluster associated with a less differentiated and less cytotoxic phenotype, as well as a lower prevalence of a subcluster associated with chemokine and cytotoxic genes. Finally, it had a major expansion of one of the CD8<sup>+</sup> Tm cells subclusters annotated as NK-like T cell and characterized by a high CCR5 mRNA expression, while the levels of CCL3, CCL4 and CCL5 mRNA were downregulated. The present findings point toward a potential link between CCL signaling and improved NK cell expansion performance, including possible markers for further investigations, and suggest future strategies to increase the final NK cell fraction and expansion based on donor-specific markers.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"28 1","pages":"Article 101994"},"PeriodicalIF":3.2,"publicationDate":"2025-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145624293","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-14DOI: 10.1016/j.jcyt.2025.10.003
Yeon Joo Lee , Yuxuan Li , Michael Scordo , Gyuri Han , Ilya Glezerman , Brian Shaffer , Sergio A. Giralt , Esperanza B. Papadopoulos , Miguel-Angel Perales , Ann A. Jakubowski , Genovefa A. Papanicolaou
Background
BK polyomavirus (BKPyV)-associated hemorrhagic cystitis occurs in 7–60% of allogeneic hematopoietic cell transplantation (HCT) recipients and may cause significant morbidity.
Objective
We conducted a pilot study to assess if early BKPyV viruria (≤day [D]+40 post-HCT) predicted risk for BKPyV hemorrhagic cystitis in the first year post-HCT.
Study Design
Adult consecutive allogeneic HCT recipients at Memorial Sloan Kettering Cancer Center from August 2019 through April 2021 were screened for urine BKPyV quantitative PCR (Eurofins Viracor, Lenexa, KS) between D-14 and D0 (pre-HCT). If the pretransplant test was negative, urine BKPyV was retested between D+20 and D+40 postengraftment. If the pre-HCT test was positive, repeat postengraftment testing was not performed. Patients with early BKPyV viruria were monitored by plasma BKPyV quantitative PCR bimonthly for the first 6 months post-HCT or 12 months post-HCT in patients with graft-versus-host disease requiring systemic corticosteroids. Patients were followed clinically for the development of BKPyV hemorrhagic cystitis in the first year post-HCT. Urine BKPyV PCR was checked per standard of care if patients developed symptoms of cystitis. BKPyV hemorrhagic cystitis was defined as BKPyV viruria with macroscopic hematuria and symptoms of cystitis. Risk factors for BKPyV hemorrhagic cystitis were examined using logistic regression models.
Results
Of 152 patients analyzed, 73 (48%) had early BKPyV viruria (33 pre-HCT and 40 post-HCT). Nine patients developed BKPyV hemorrhagic cystitis before D+40, and 7 patients after D+40. Among the 79 patients without early BKPyV viruria, none developed BKPyV hemorrhagic cystitis. The cumulative incidence of BKPyV hemorrhagic cystitis by 1-year post-HCT was 10% (95% confidence interval: 0.051–0.146). Cut point analysis demonstrated that BKPyV urine viral load ≥6.35 × 106 copies/mL was the optimal discriminator for BKPyV hemorrhagic cystitis at 1-year post-transplant (odds ratio: 7.59, 95% confidence interval: 2.27, 27.2; P = .001).
Conclusions
Nearly half of HCT patients had BKPyV viruria by D+40 post-HCT. Early BKPyV viruria ≥6.35 × 106 copies/mL may help identify patients at risk for BKPyV hemorrhagic cystitis. Clinical trials of BKV therapeutics may use early BKPyV screening to identify patients at high-risk for BKPyV hemorrhagic cystitis.
{"title":"Pilot study of early BK polyomavirus in the urine and risk of hemorrhagic cystitis after allogeneic hematopoietic cell transplantation","authors":"Yeon Joo Lee , Yuxuan Li , Michael Scordo , Gyuri Han , Ilya Glezerman , Brian Shaffer , Sergio A. Giralt , Esperanza B. Papadopoulos , Miguel-Angel Perales , Ann A. Jakubowski , Genovefa A. Papanicolaou","doi":"10.1016/j.jcyt.2025.10.003","DOIUrl":"10.1016/j.jcyt.2025.10.003","url":null,"abstract":"<div><h3>Background</h3><div>BK polyomavirus (BKPyV)-associated hemorrhagic cystitis occurs in 7–60% of allogeneic hematopoietic cell transplantation (HCT) recipients and may cause significant morbidity.</div></div><div><h3>Objective</h3><div>We conducted a pilot study to assess if early BKPyV viruria (≤day [D]+40 post-HCT) predicted risk for BKPyV hemorrhagic cystitis in the first year post-HCT.</div></div><div><h3>Study Design</h3><div>Adult consecutive allogeneic HCT recipients at Memorial Sloan Kettering Cancer Center from August 2019 through April 2021 were screened for urine BKPyV quantitative PCR (Eurofins Viracor, Lenexa, KS) between D-14 and D0 (pre-HCT). If the pretransplant test was negative, urine BKPyV was retested between D+20 and D+40 postengraftment. If the pre-HCT test was positive, repeat postengraftment testing was not performed. Patients with early BKPyV viruria were monitored by plasma BKPyV quantitative PCR bimonthly for the first 6 months post-HCT or 12 months post-HCT in patients with graft-versus-host disease requiring systemic corticosteroids. Patients were followed clinically for the development of BKPyV hemorrhagic cystitis in the first year post-HCT. Urine BKPyV PCR was checked per standard of care if patients developed symptoms of cystitis. BKPyV hemorrhagic cystitis was defined as BKPyV viruria with macroscopic hematuria and symptoms of cystitis. Risk factors for BKPyV hemorrhagic cystitis were examined using logistic regression models.</div></div><div><h3>Results</h3><div>Of 152 patients analyzed, 73 (48%) had early BKPyV viruria (33 pre-HCT and 40 post-HCT). Nine patients developed BKPyV hemorrhagic cystitis before D+40, and 7 patients after D+40. Among the 79 patients without early BKPyV viruria, none developed BKPyV hemorrhagic cystitis. The cumulative incidence of BKPyV hemorrhagic cystitis by 1-year post-HCT was 10% (95% confidence interval: 0.051–0.146). Cut point analysis demonstrated that BKPyV urine viral load ≥6.35 × 10<sup>6</sup> copies/mL was the optimal discriminator for BKPyV hemorrhagic cystitis at 1-year post-transplant (odds ratio: 7.59, 95% confidence interval: 2.27, 27.2; <em>P</em> = .001).</div></div><div><h3>Conclusions</h3><div>Nearly half of HCT patients had BKPyV viruria by D+40 post-HCT. Early BKPyV viruria ≥6.35 × 10<sup>6</sup> copies/mL may help identify patients at risk for BKPyV hemorrhagic cystitis. Clinical trials of BKV therapeutics may use early BKPyV screening to identify patients at high-risk for BKPyV hemorrhagic cystitis.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"28 3","pages":"Article 101993"},"PeriodicalIF":3.2,"publicationDate":"2025-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145446497","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-11DOI: 10.1016/j.jcyt.2025.10.001
Utsav Jetley, Ishina Balwani , Palak Sharma, Ian C. Miller , Allie Luther, Ivy Dutta, Nithila Saravanan, Surbhi Goel, Qingzhan Zhang, Boning Zhang , Ozgun Kilic , Biao Liu, Bo W. Han, Dai Liu, Birgit Schultes, Aaron Prodeus, Yong Zhang
Background aims
Autologous T-cell therapies have shown profound clinical responses; however, their widespread use has been limited primarily as the result of their individualized manufacturing requirements.
Methods
To develop a persistent “off-the-shelf” allogeneic (Allo) approach, a multiplex Nme2Cas9-based cytosine base editor was deployed to knockout select human leukocyte antigens (HLA) class I and II alleles (HLA-A, HLA-B and the class II transactivator [CIITA]) while retaining HLA-C to protect from natural killer (NK) cell rejection.
Results and Conclusion
Matching the residual HLA-C allele from homozygous donors to the host prevented rejection of the donor T cells by allogeneic host T and NK cells. Site-specific integration of a tumor-specific CAR or TCR into the TRAC locus using SpyCas9 nuclease and an adeno-associated virus template allowed for a high localized insertion rate while simultaneously removing the endogenous TCR and preventing graft-versus-host disease. Using an optimized T-cell engineering process involving orthogonal CRISPR/Cas9 cleavage and base editors coupled with lipid nanoparticle delivery, we achieved efficient production of Allo-CAR T cells with high editing rates and cell expansion in a scalable manner. These allogeneic T cells demonstrated comparable functional activity to their autologous counterparts in preclinical assays. Moreover, this gene-editing approach significantly minimized the occurrence of chromosomal aberrations. This promising allogeneic approach also has been applied to induced pluripotent stem cells (iPSCs) with triple edits targeting HLA-A, HLA-B and CIITA (TKO). Pancreatic progenitor cells or cardiomyocytes derived from TKO iPSCs were protected from host peripheral blood mononuclear cell−mediated rejection when matched for HLA-C, suggesting potential applications in regenerative medicine applications.
{"title":"A differentiated and durable allogeneic strategy applicable to cell therapies","authors":"Utsav Jetley, Ishina Balwani , Palak Sharma, Ian C. Miller , Allie Luther, Ivy Dutta, Nithila Saravanan, Surbhi Goel, Qingzhan Zhang, Boning Zhang , Ozgun Kilic , Biao Liu, Bo W. Han, Dai Liu, Birgit Schultes, Aaron Prodeus, Yong Zhang","doi":"10.1016/j.jcyt.2025.10.001","DOIUrl":"10.1016/j.jcyt.2025.10.001","url":null,"abstract":"<div><h3>Background aims</h3><div>Autologous T-cell therapies have shown profound clinical responses; however, their widespread use has been limited primarily as the result of their individualized manufacturing requirements.</div></div><div><h3>Methods</h3><div>To develop a persistent “off-the-shelf” allogeneic (Allo) approach, a multiplex Nme2Cas9-based cytosine base editor was deployed to knockout select human leukocyte antigens (HLA) class I and II alleles (<em>HLA-A, HLA-B</em> and the class II transactivator [<em>CIITA</em>]) while retaining HLA-C to protect from natural killer (NK) cell rejection.</div></div><div><h3>Results and Conclusion</h3><div>Matching the residual HLA-C allele from homozygous donors to the host prevented rejection of the donor T cells by allogeneic host T and NK cells. Site-specific integration of a tumor-specific CAR or TCR into the <em>TRAC</em> locus using SpyCas9 nuclease and an adeno-associated virus template allowed for a high localized insertion rate while simultaneously removing the endogenous TCR and preventing graft-versus-host disease. Using an optimized T-cell engineering process involving orthogonal CRISPR/Cas9 cleavage and base editors coupled with lipid nanoparticle delivery, we achieved efficient production of Allo-CAR T cells with high editing rates and cell expansion in a scalable manner. These allogeneic T cells demonstrated comparable functional activity to their autologous counterparts in preclinical assays. Moreover, this gene-editing approach significantly minimized the occurrence of chromosomal aberrations. This promising allogeneic approach also has been applied to induced pluripotent stem cells (iPSCs) with triple edits targeting <em>HLA-A, HLA-B</em> and <em>CIITA</em> (TKO)<em>.</em> Pancreatic progenitor cells or cardiomyocytes derived from TKO iPSCs were protected from host peripheral blood mononuclear cell−mediated rejection when matched for HLA-C, suggesting potential applications in regenerative medicine applications<em>.</em></div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"28 3","pages":"Article 101991"},"PeriodicalIF":3.2,"publicationDate":"2025-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145460320","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-02DOI: 10.1016/j.jcyt.2025.09.013
Alexandra Dreyzin , Anne Marijn Kramer , Bonnie Yates , Hao-Wei Wang , Bita Sahaf , Constance Yuan , Dorota Klysz , Ramya Tunuguntla , Zachary Ehlinger , Skyler Reitberg , Serifat Adebola , Angela Su , Risa Ebina-Shibuya , Dongya Jia , Sooraj Achar , Sunita Patil , Kathryn Martin , Nikeshan Jeyakumar , Kara L Davis , Terry Fry , Nirali N Shah
As use of chimeric antigen receptor (CAR) T-cells continues to grow, there is increasing interest in utilizing automated manufacturing systems as a mechanism to support decentralized manufacturing and increase access. However, most FDA approved CAR T-cell therapies are manufactured using traditional bag culture methodologies. Thus, understanding how different manufacturing platforms may impact outcomes is imperative. With parallel trials of CD22 CAR T-cells conducted in patients with B-cell acute lymphoblastic leukemia using a uniform vector but two different manufacturing strategies – either bag-culture (BC) or Prodigy – we were able to compare outcomes. Across 57 patients, 41 received BC cells and 16 received Prodigy-based cells. No significant differences in response rates or incidence of CAR-associated toxicities were observed between cohorts, although the BC cohort had slightly higher rates of severe CRS and IEC-HS. Peak ferritin and C-reactive protein levels were higher in the BC cohort. CAR T-cell expansion was similar, except for patients who had extramedullary disease with low bone marrow disease burden (n = 6 from each group), for whom BC-manufactured cells had greater expansion. In summary, while efficacy across both platforms was comparable, lower inflammatory markers in those who received Prodigy manufactured CAR T-cells suggest changes in the infusion product.
{"title":"Outcomes following CD22 CAR T-cells in B-ALL: a tale of two manufacturing strategies","authors":"Alexandra Dreyzin , Anne Marijn Kramer , Bonnie Yates , Hao-Wei Wang , Bita Sahaf , Constance Yuan , Dorota Klysz , Ramya Tunuguntla , Zachary Ehlinger , Skyler Reitberg , Serifat Adebola , Angela Su , Risa Ebina-Shibuya , Dongya Jia , Sooraj Achar , Sunita Patil , Kathryn Martin , Nikeshan Jeyakumar , Kara L Davis , Terry Fry , Nirali N Shah","doi":"10.1016/j.jcyt.2025.09.013","DOIUrl":"10.1016/j.jcyt.2025.09.013","url":null,"abstract":"<div><div>As use of chimeric antigen receptor (CAR) T-cells continues to grow, there is increasing interest in utilizing automated manufacturing systems as a mechanism to support decentralized manufacturing and increase access. However, most FDA approved CAR T-cell therapies are manufactured using traditional bag culture methodologies. Thus, understanding how different manufacturing platforms may impact outcomes is imperative. With parallel trials of CD22 CAR T-cells conducted in patients with B-cell acute lymphoblastic leukemia using a uniform vector but two different manufacturing strategies – either bag-culture (BC) or Prodigy – we were able to compare outcomes. Across 57 patients, 41 received BC cells and 16 received Prodigy-based cells. No significant differences in response rates or incidence of CAR-associated toxicities were observed between cohorts, although the BC cohort had slightly higher rates of severe CRS and IEC-HS. Peak ferritin and C-reactive protein levels were higher in the BC cohort. CAR T-cell expansion was similar, except for patients who had extramedullary disease with low bone marrow disease burden (n = 6 from each group), for whom BC-manufactured cells had greater expansion. In summary, while efficacy across both platforms was comparable, lower inflammatory markers in those who received Prodigy manufactured CAR T-cells suggest changes in the infusion product.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"28 3","pages":"Article 101990"},"PeriodicalIF":3.2,"publicationDate":"2025-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145446512","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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