Haploidentical Natural Killer Cells Research Articles

Safety and Virologic Impact of Haploidentical NK Cells Plus Interleukin 2 or N-803 in HIV Infection.

Natural killer (NK) cells are dysfunctional in chronic human immunodeficiency virus (HIV) infection as they are not able to clear virus. We hypothesized that an infusion of NK cells, supported by interleukin 2 (IL-2) or IL-15, could decrease virus-producing cells in the lymphatic tissues. We conducted a phase 1 pilot study in 6 persons with HIV (PWH), where a single infusion of haploidentical related donor NK cells was given plus either IL-2 or N-803 (an IL-15 superagonist). The approach was well tolerated with no unexpected adverse events. We did not pretreat recipients with cyclophosphamide or fludarabine to "make immunologic space," reasoning that PWH on stable antiretroviral treatment remain T-cell depleted in lymphatic tissues. We found donor cells remained detectable in blood for up to 8 days (similar to what is seen in cancer pretreatment with lymphodepleting chemotherapy) and in the lymph nodes and rectum up to 28 days. There was a moderate decrease in the frequency of viral RNA-positive cells in lymph nodes. There was a moderate decrease in HIV-producing cells in lymph nodes. Further studies are warranted to determine the impact of healthy NK cells on HIV reservoirs and if restoring NK-cell function could be part of an HIV cure strategy. Clinical Trials Registration. NCT03346499 and NCT03899480.

A Phase I, Single-Arm, Open Label, Dose Escalation, Multicenter Study of Off-the-Shelf Natural Killer (NK) Cells (SAR445419) in Patients with Relapsed/Refractory Acute Myeloid Leukemia (R/R AML)

Background and Significance: SAR445419 (SAR'419) is an allogeneic, off-the-shelf, donor-derived, natural killer (NK) cell investigational immunotherapy. NK cells in SAR'419 are expanded ex vivo using PM21 particles expressing membrane-bound interleukin-21 and 4-1BB ligand from the plasma membranes of feeder cells (Ciurea et al, Leukemia, 2022). Ex vivo, PM21 expansion allows for large numbers of hyperactive NK cells to be generated and administered, thereby increasing therapeutic potential to patients. It is hypothesized that mature, fully functional NK cells expanded ex vivo will have superior anti-tumor activity, favorable safety profile, and can induce remissions, enabling patients to receive curative allogeneic hematopoietic stem cell transplantation (HSCT), thereby improving outcomes in patients with R/R AML. Two phase 1/2 dose-escalation and expansion studies (NCT01787474, NCT02809092) of ex vivo expanded haploidentical donor NK cells infused as 6 doses over a course of 2 weeks, after fludarabine and cytarabine chemotherapy, demonstrated this approach to be feasible with favorable safety profile in patients with R/R AML. This study aims to determine the candidate dose(s), evaluate safety, tolerability, and preliminary anti-tumor activity of SAR'419 administered after fludarabine and cytarabine chemotherapy for adult patients with R/R AML. Study Design and Methods: This is a phase 1, single-arm, dose escalation study (NCT05712278). The primary objective is to determine the candidate dose(s) based on the evaluation of dose-limiting toxicities (DLTs). The DLT evaluation period is from start of chemotherapy (Day -6) until 28 days after the first administration of SAR'419, starting at the lowest dose. Eligible patients will be administered fludarabine, 30 mg/m 2/day and cytarabine, 2 g/m 2/day (Day -6 to Day -2) for 5 days followed by 6 doses of SAR'419 (intravenous) given thrice weekly over 2 weeks beginning on Day 1. Approximately 12 DLT-evaluable patients will be enrolled. The treatment period will start from the first administration of chemotherapy (Day -6) until end of treatment on Day 56, and subsequently patients will have follow-up visits every 2 months. Dose escalation will be guided by the modified toxicity probability interval 2 design. Key secondary objectives include characterization of overall safety, tolerability, HSCT rate, impact of SAR'419 on infection, and preliminary anti-leukemic activity of SAR'419 (as per international working group criteria). Biomarkers in whole blood (including serum and plasma) and bone marrow (aspirates-liquid, and biopsy-solid) will be evaluated to assess the impact of SAR'419 treatment. Key inclusion criteria are adult participants with confirmed diagnosis of R/R AML according to World Health Organization classification, including participants with relapsed AML in the bone marrow after allogeneic HSCT (including those who have received donor lymphocyte infusions); with or without concomitant or with isolated central nervous system or extramedullary disease who have received at least 1 prior line of therapy for AML. Key exclusion criteria are participants with a second primary malignancy requiring active therapy (adjuvant hormonal therapy is allowed); with known acquired immunodeficiency syndrome or human immunodeficiency virus disease requiring antiretroviral treatment or having active hepatitis B or C infection, or symptomatic severe acute respiratory syndrome coronavirus 2 infection; and who are receiving >10 mg/day of oral prednisone or equivalent. In the safety analyses, all DLT modeling will be performed on the DLT-evaluable population or those who experienced a DLT. All other safety analyses will be performed on the safety population using descriptive statistics. This study is actively recruiting at this time and is being conducted in the United States.

CTLA-4 Blockade Results in the Enrichment of Proliferative CD56 dimCD16 + NK Cells Following Infusion of Haploidentical Donor Memory-like Natural Killer Cells Plus IL-15 Superagonist in a Phase 1 Trial

Introduction Regulatory T cells (Tregs) blunt anti-tumor responses of major immune effector cells including CD8 T cells and natural killer (NK) cells. Tregs constitutively express CTLA-4 and thus the use of ipilimumab, a CTLA-4 blocker can potentially overcome the Treg-mediated immune suppression. We (and others) have shown that a brief activation of conventional NK cells with the cytokine combination IL-12, IL-15, and IL-18 generates cytokine-induced memory-like (CIML) NK cells which exhibit enhanced anti-tumor responses, have a longer half-life (up to 90 vs ~12 days of conventional NK cells), and show promising activity in several early phase clinical trials. N-803 is an IL-15 super-agonist with a long half-life that preferentially activates and expands NK and CD8+ T cells without affecting Tregs, making it an ideal cytokine for combining with adoptive immunotherapy. In the current study, we hypothesized the use of ipilimumab (IPI) will abrogate Treg-mediated inhibition and thus allow enhanced proliferation, activation, and anti-tumor responses of the adoptively transferred CIML NK cells.To address this, we conducted a phase 1 trial of HLA-haploidentical CIML NK cells in combination with IPI and N-803 in patients with advanced head and neck cancer (HNC). Using flow cytometry and CITE-Seq on the banked peripheral blood samples, we performed an in-depth characterization of NK cells expanded following IPI pre-exposure and compared it to those who did not receive IPI. Methods We enrolled 10 patients with refractory HNC regardless of human papillomavirus (HPV) status who had prior platinum and immunotherapy (NCT04290546). All patients received lymphodepletion with fludarabine (25 mg/m 2 x 5 days) and cyclophosphamide (60 mg/kg x 2 days) during days -6 to -2 prior to haploidentical CIML NK cell infusion on day 0 (5-10 x 10 6 cells/kg=dose level 0) followed by N-803 (15 mcg/kg subcutaneously) starting on day +1 every 21-days for 4-doses. In cohort 2, patients received IPI (3 mg/kg) on day -7, no ipilimumab was given to the first 6 patients treated on the lead-in safety cohort 1. Flow cytometry using a custom NK cell panel was performed on samples collected on days +7, +14, +21, +28, +42, and +60 after CIML NK cell infusion. FlowSOM clustering was performed on NK cell markers. UMAP clustering was applied to CITE sequencing data acquired from samples collected on day +7 and day +28, and well-defined markers genes for each cluster were used to identify cell populations. NK cell function (cytotoxicity, IFNg, and CD107a) was assessed using flow-cytometry based assays. Results The median age of the patients was 58, 90% were men and 80% had oropharyngeal primaries. Mild CRS (grade 2 or lower) was observed in 60% of patients. No neurotoxicity or graft-versus-host disease was observed. Maximal expansion of NK cells occurred on day +7 following CIML NK cell infusion in 9/10 of patients. The single patient without NK cell expansion was found to have a donor-specific anti-HLA antibody. Day +7 NK cells exhibited upregulation of IFNg expression following cytokine restimulation as compared to resting NK cells. Tumor regression was observed in 50% of treated patients at day +30 based on RECIST v1.1 criteria. Patients who had tumor regression had an expansion of CD56 dimCD16 + NK cell clusters by day +14. IPI treated patients were noted to have a relative enrichment of CD16 + NK cell metaclusters at day +14 compared to no IPI patients (Figure 1A). At the time of maximal NK cell expansion, CITEseq defined NK cell clusters were dominant in both IPI and no IPI patients, but the proliferative gene sets were enriched in the IPI treated patient (Figure 1B). IPI exposure had no effect on the distribution of regulatory T cells or CD8 + T cell populations at day +28 compared to the no IPI patients. Conclusions The use of donor-derived CIML NK cells is safe and associated with tumor regression in patients with advanced HNC. The use of IPI was associated with the preferential expansion of CD16 + NK cell clusters that had enrichment of proliferative gene sets. Tumor regression was associated with CD56 dimCD16 + NK cell expansion. Further work is required to elucidate the mechanism by which IPI exposure is associated with the expansion of specific subsets of CIML NK cells.

Intra-lesion injection of activated Natural Killer (NK) cells in recurrent malignant brain tumors

Despite multi-modal therapies for patients with malignant brain tumors, their median survival is < 2 years. Recently, NK cells have provided cancer immune surveillance through their direct natural cytotoxicity and by modulating dendritic cells to enhance the presentation of tumor antigens and regulate T-cell-mediated antitumor responses. However, the success of this treatment modality in brain tumors is unclear. The main reasons are; the brain tumor microenvironment, the NK cell preparations and administration, and the donor selection. Our previous study showed that intracranial injection of activated haploidentical NK cells resulted in the eradication of glioblastoma tumor mass in the animal model without any evidence of tumor recurrence. Therefore, in the present study, we evaluated the safety of intra-surgical cavity or intra cerebrospinal fluid (CSF) Injectionofex vivoactivated haploidentical NK cells in six patients with recurrent glioblastoma multiform (GBM) and malignant brain tumors resistance to chemo/radiotherapy. Our results indicated that activated haploidentical NK cells express activator and inhibitor markers and can kill the tumor cells. However, their cytotoxic potential on patient-derived GBM (PD-GBM) was more than that of its cell line. Also, their infusion increased the overall disease control rate by about 33.3%, with a mean survival of 400 days. Moreover, we showed that local administration of the activated haploidentical NK cells in malignant brain tumors is safe, feasible, tolerated at higher doses, and cost-effective.

Haploidentical Natural Killer Cell Therapy as an Adjunct to Stem Cell Transplantation for Treatment of Refractory Acute Myeloid Leukemia

Refractory acute myeloid leukemia (AML), defined as failure of two cycles of induction therapy at diagnosis or of one cycle at relapse, represents a subgroup with poor outcomes. Haploidentical natural killer cell (NK) therapy is a strategy that is being explored in refractory malignancies. Historically, at our center, patients with refractory AML have been treated with cytoreductive therapy (fludarabine + cytosine + granulocyte colony-stimulating factor ± idarubicin or mitoxantrone + etoposide) followed by 1-week rest and then reduced-intensity transplant with fludarabine + melphalan. We used the same backbone for this trial (CTRI/2019/02/017505) with the addition of CD56-positive cells from a family donor infused 1 day after the completion of chemotherapy. CD56-positive selection was done using a CliniMACS Prodigy system (Miltenyi Biotec, Bergisch Gladbach, Germany) followed by overnight incubation in autologous plasma with 2 micromolar arsenic trioxide and 500 U/mL of interleukin-2. From February 2019, 14 patients with a median age of 29 years (interquartile range [IQR]: 16.5–38.5) were enrolled in this trial. Six were females. Six had primary refractory AML while eight had relapsed refractory AML. The median CD56-cell dose infused was 46.16 × 106/kg (IQR: 25.06–70.36). One patient withdrew consent after NK cell infusion. Of the 13 patients who proceeded to transplant, five died of immediate post-transplant complications while two did not engraft but were in morphologic leukemia-free state (both subsequently died of infective complications after the second transplant). Of the remaining six patients who engrafted and survived beyond 1 month of the transplant, two developed disease relapse and died. The remaining four patients are alive and relapse free at the last follow-up (mean follow-up duration of surviving patients is 24 months). The 2-year estimated overall survival for the cohort was 28.6% ± 12.1% while the treatment-related mortality (TRM) with this approach was 38.5% ± 13.5%. Haploidentical NK cell therapy as an adjunct to transplant is safe and needs further exploration in patients with AML. For refractory AML, post-transplant NK infusion and strategies to reduce TRM while using pre-transplant NK infusion merit exploration.

A Novel Dual-Antigen Targeting Approach Enables Off-the-Shelf CAR NK Cells to Effectively Recognize and Eliminate the Heterogenous Population Associated with AML

While acute myeloid leukemia (AML) treatment has improved, relapse is still common, even after reduced intensity conditioning allogeneic transplantation. Natural killer (NK) cells are a promising allogeneic cell immunotherapy without the risk of graft-versus-host disease or cytokine release syndrome. We and others have reported that haploidentical NK cells can safely result in 30-50% complete remissions in advanced AML patients when given with high-dose lymphodepleting chemotherapy and exogenous cytokine support. However, while NK cell natural cytotoxicity has a clinical signal in AML, increased specificity and multi-dosing are likely to enhance the efficacy and durability of this treatment strategy. A number of stress ligands, including the MHC-I polypeptide-related sequence A and B (MICA/B), bind to the NK cell activating receptor NKG2D and initiate NK cell killing of transformed and infected cells, and several groups have shown promising results in clinical trials by developing NK cells that carry a NKG2D chimeric antigen receptor (CAR). However, the distal α1 and α2 domains of MICA/B that are recognized by NKG2D can be proteolytically cleaved from the surface of the tumor by metalloproteases, allowing for escape from NK cell-mediated detection and elimination. Wucherpfennig and colleagues have described that the membrane proximal α3 domain remains on the cell surface and may be a potential target for immunotherapy across many cancers, including AML. However, since AML is heterogenous, we hypothesized that clinical success would require dual-targeting and selected CD33 as the ideal complementary targeting approach since CD33 is a validated marker expressed on greater than 80% of AML blasts. To this end, our lab has developed a Tri-specific Killer Engager (TriKE) capable of agonistically ligating the CD16 Fc receptor to CD33 found on AML along with IL-15 co-stimulation to further activate the NK cell response. Here, we demonstrate the efficacy of a multiplexed-gene edited induced pluripotent stem-cell (iPSC) derived NK cell (iNK) product that expresses a high-affinity, non-cleavable version of CD16 (hnCD16), a membrane-bound IL-15 fusion receptor, CD38 knockout to enhance metabolic fitness, and a CAR against the α3 domain of MICA/B (α3 MICA/B) to drive a potent response against AML alone and when combined with anti-CD33 TriKE. In the initial study, the iNK cell backbone (iNK cells without the α3 MICA/B CAR) induced potent activity against the AML cell line HL60, and displayed further enhancement of activity with the addition of anti-CD33 TriKE (GTB-3650), representing combined effects of natural cytotoxicity and antibody-dependent cellular cytotoxicity. To show specificity for α3 MICA/B targeting, the AML cell line THP-1 was stained for the presence of MICA/B by flow cytometry using the 6D4 clone that recognizes the α1/2 domains of MICA/B or the 7C6 clone that uniquely recognizes the α3 domain +/- proteolytic cleavage with trypsin (Fig. 1A-B). Significant expression of the α3 domain was observed on THP-1 cells using the 7C6 antibody, which was highly expressed even after protease treatment. On the other hand, the α1/2 domains of MICA/B were detectable at lower levels using the 6D4 antibody and were undetectable after protease treatment (Fig. 1A-B). To assess a dual targeting approach, THP-1 cells were used as targets in live imaging functional assays under standard conditions (Effector:Target [E:T] 2:1). The iNK cell backbone had modest natural cytotoxicity at 3 hours that was significantly enhanced by the addition of α3 MICA/B CAR. The effect was further improved with more rapid killing kinetics when combined with the anti-CD33 TriKE (Fig. 1C). To evaluate the efficacy of dual-targeting in a more stringent physiologic manner, we mimicked "stress” conditions by minimizing the E:T ratio to 0.25:1. While all effector conditions induced immediate killing in 4 hours at the low E:T ratio, sustained tumor control was only observed with dual-antigen targeting (Fig. 1D). Studies with primary AML targets, +/- preincubation of decitabine and all-trans retinoic acid, known to upregulate NKG2D ligands in AML, are in progress and will be discussed. In summary, dual-targeting strategies using off-the-shelf CAR NK cells targeting α3 MICA/B in combination with antigen-specific TriKE targeting CD33 represent an ideal clinical strategy to enhance efficacy and durability of treatment in advanced AML. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Abstract CT540: A phase 1 trial of cytokine-induced memory-like (CIML) natural killer (NK) cell therapy with IL-15 superagonist in advanced head and neck cancer: Part 1 results

Abstract Purpose: Outcomes for patients with recurrent, incurable or metastatic (R/M) squamous cell carcinoma of the head and neck (SCCHN) refractory to platinum and immunotherapy are poor. Cellular therapies are emerging treatments with potential utility in epithelial cancers. This proof-of-concept trial investigates an allogeneic cytokine-induced, memory-like (CIML) NK cell infusion with IL-15 superagonist (sa) after lymphodepleting (LD) chemotherapy in advanced SCCHN. Patients and methods: This phase 1 single-center trial enrolled patients (pts) with R/M SCCHN regardless of human papillomavirus (HPV) status who had prior platinum and immunotherapy. Pts received LD fludarabine (25 mg/m2) and cyclophosphamide (60 mg/m2/kg) on days -6 to -2 prior to haploidentical CIML NK cell infusion on day 0 (5-10 x 106 viable cells/kg=dose level 0) followed by N-803 (IL-15sa, 15 mcg/kg subcutaneously) starting on day +1 every 21-days for 4-doses. Part 1 treated 3 pts at dose level 0; &amp;lt;2 DLTs triggered an additional 3 pts. Part 2 will treat an additional 6 pts with lead-in ipilimumab (day -7). Primary objective: safety, maximum tolerated dose of CIML NK cells. Secondary objectives: objective response rate, progression-free survival (PFS), overall survival (OS), and phenotypic expansion and function of adoptively transferred NK cells. Results: From 9/8/20 to 9/7/21, 6 pts enrolled to Part 1. One DLT was observed at dose level 0. Among 6 pts, median age: 59; 5/6 (83%) were men; 5/6 (83%) had oropharyngeal primaries (4 HPV+) with a median 6 prior lines of therapy for R/M disease (range: 4-8). R/M disease sites: lung, bone, skin, liver. 5/6 (83%) had offspring donors. Grade (G) 3-4 hematologic adverse events were common (6/6, 100%). One patient died of G5 febrile neutropenia and infection. Median days hospitalized: 14 (range: 9-37). Mild cytokine release syndrome was observed in 5/6 (83%) (median peak ferritin: 2248, CRP: 168); 3/5 required anti-IL6 therapy; no neurotoxicity was noted. There were no dose adjustments or discontinuation of therapy. One (17%) partial response (PR) was observed lasting 6.5 months; 4 (67%) pts had stable disease (SD), and 1 (17%) had progression. Tumor regression was observed in 3/6 (50%) pts at day +30. At a median follow-up of 9.5 months, median PFS: 3.4 months (95%CI 2.6-6.5); median OS: 4.7 months (95%CI 3.4-11.8). CIML NK cells showed large expansion in the peripheral blood (PB) at day +7 in all pts; mean increase: 66% (6-fold; standard deviation [SD] 10.5), to constitute 80% (SD 12.1) of PB lymphocytes. In pts with tumor regression at day +30 compared to those without, the % of PB NK cells remained high at day +28 (mean: 78 vs. 11%). PB NK cells remained &amp;gt;50% at day +42 in the pt with a PR. Conclusion: Allogeneic CIML NK cells can induce tumor regression associated with persistent CIML NK cell expansion among advanced SCCHN pts. In Part 1 we demonstrate safety and feasibility with the expected risks of LD conditioning. These findings have important implications for the development of cellular therapies in solid tumors. Citation Format: Glenn J. Hanna, Kimberly Coleman, Grace Birch, Robert A. Redd, Alejandro Alonso, Samantha Bednarz, Heather Daley, Diego E. Hernandez Rodriguez, Kit L. Shaw, Robert I. Haddad, Ravindra Uppaluri, Jerome Ritz, Sarah Nikiforow, Robert J. Soiffer, Rizwan Romee. A phase 1 trial of cytokine-induced memory-like (CIML) natural killer (NK) cell therapy with IL-15 superagonist in advanced head and neck cancer: Part 1 results [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr CT540.

284 - Haploidentical Natural Killer Cell Therapy As an Adjunct to Stem Cell Transplantation for Refractory Acute Myeloid Leukemia

284 - Haploidentical Natural Killer Cell Therapy As an Adjunct to Stem Cell Transplantation for Refractory Acute Myeloid Leukemia

Sub-myeloablative Second Transplantations with Haploidentical Donors and Post-Transplant Cyclophosphamide have limited Anti-Leukemic Effects in Pediatric Patients

Pediatric patients with high-risk hematologic malignancies who experience relapse after a prior allogeneic hematopoietic cell transplant (HCT) have an exceedingly poor prognosis. A second allogeneic HCT offers the potential for long-term cure but carries high risks of both subsequent relapse and HCT-related morbidity and mortality. Using haploidentical donors for HCT (haploHCT) can expand the donor pool and potentially enhance the graft-versus-leukemia effect but is accompanied by a risk of graft-versus-host disease (GVHD). The goal of this protocol was to intensify the antileukemia effect of haploHCT for pediatric patients with hematologic malignancies that relapsed after prior allogeneic HCT, while limiting regimen-associated toxicities. This phase II clinical trial evaluated a sub-myeloablative preparative regimen consisting of anti-thymocyte globulin, clofarabine, cytarabine, busulfan, and cyclophosphamide, in combination with plerixafor to sensitize leukemic blasts. Participants received a mobilized peripheral blood unmanipulated haploidentical donor graft with one dose of post-transplant cyclophosphamide as GVHD prophylaxis, followed by natural killer (NK) cell addback. Here we report the clinical outcomes and immune reconstitution of 17 participants treated on the study and 5 additional patients treated on similar single-patient treatment plans. Of the 22 participants analyzed, 12 (55%) had active disease at the time of HCT. The regimen provided robust immune reconstitution, with 21 participants (95%) experiencing neutrophil engraftment at a median of 14 days after HCT. In this high-risk population, the overall survival was 45% (95% confidence interval [CI], 24%-64%), with a 12-month event-free survival of 31% (95% CI, 14%-51%) and cumulative incidence of relapse at 12 months of 50% (95% CI, 27%-69%). Four participants (18%) remain in remission at >5 years follow-up. Expected HCT-related organ-specific toxicities were observed, and 13 participants (59%) experienced acute or chronic GVHD. This intensified but sub-myeloablative regimen, followed by a high-dose unmanipulated haploidentical graft, post-transplantation cyclophosphamide, and NK cell infusion, resulted in adequate immune reconstitution but failed to overcome the elevated risks of relapse and treatment-related morbidity in this high-risk population.

Haploidentical Natural Killer Cell Therapy As an Adjunct to Stem Cell Transplantation for Refractory Acute Myeloid Leukemia

Refractory acute myeloid leukemia (AML), defined as failure of 2 cycles of induction therapy at diagnosis or of 1 cycle at relapse, represents a subgroup with poor clinical outcomes. In our transplant cohort, the 5-year overall survival in this subgroup was 16% (Ganapule at al. JGO 2017). Haploidentical natural killer cell (NK) therapy is a strategy that is being explored in refractory malignancies. Our in-vitro and animal model data suggest that exposure to arsenic trioxide (ATO) results in enhanced NK cytotoxicity (Alex AA et al. Front. Immunol 2018). Historically, at our center, patients with refractory AML have been treated with cytoreductive therapy (FLAG ± idarubicin or mitoxantrone + etoposide for 3 to 5 days) followed by 1-week rest and then a reduced-intensity transplant with fludarabine + melphalan conditioning while in peak cytopenia. From February 2019, we initiated a phase II single arm clinical trial (CTRI/2019/02/017505) enrolling patients with refractory AML planned for a stem cell transplant in peak cytopenia. Patients received CD56-positive cells from an HLA haploidentical related family donor (other than the stem cell donor, wherever feasible) following cytoreductive chemotherapy. The NK cell donor preference strategy included presence of KIR ligand mismatch, greater number of KIR B motifs (or the B score), lower donor age, and negative donor specific antibodies tested using flowcytometry crossmatch (Figure 1a). CD56-positive selection was done using CliniMACS prodigy system. This was followed by overnight incubation of the CD56 positive cells in autologous plasma with 2 micromolar ATO and 500 U/mL of interleukin-2. The CD56 positive cells were then infused to the patient 1-day after the completion of cytoreductive chemotherapy. This was followed by a reduced intensity stem cell transplant (Figure 1b). The primary outcome variable was 1-year relapse free survival. From February 2019, 14 patients with median age 28 years (IQR: 15.75-31.5) were enrolled in this trial. Six were females. Six had primary-refractory AML while 8 had relapsed-refractory AML. The cytoreductive chemotherapy was FLAG ± idarubicin (n=7), Mitoxantrone + Etoposide (n=6) and GCLAC (n=1). The median blast percentage on flowcytometry MRD testing prior to NK infusion was 15.9% (IQR: 9.1%-54.5%) (n=11). The median B score for the NK cell donors was 2 (IQR: 1-3). The median age of the NK cell donor was 43 years (IQR: 36-49.5). KIR ligand mismatch with the patient was noted in 2 donors. The median CD56-cell dose infused was 46.16 x 10 6/kg (IQR: 25.06-70.36) (Figure 1c). Pre-defined release criteria, including sterile cultures, and endotoxin negativity were met in all cases. There was no infusion related toxicity. The median blast percentage on flowcytometry MRD testing done following NK cell infusion was 11.9% (IQR: 4.9%-47.6%) (n=8). One patient withdrew consent after NK cell infusion and did not undergo transplant. For the remaining 13 patients, the stem cell donor was HLA matched (n=4), HLA 9/10 matched (n=1) or HLA haplomatched (n=8). The median CD34 cell dose infused was 10 x 10 6/kg (IQR:7.51-11.6). Five (38.5%) patients died of immediate post-transplant complications (sepsis (n=3) on days 1, 2 and 28, cerebral venous sinus thrombosis (n=1) on day 1 in a patient treated with hormonal contraceptives for menorrhagia, and veno-occlusive disease (n=1) on day 15 in a patient undergoing a second transplant) while 2 (15.4%) did not engraft (both subsequently died of infective complications following engraftment post-second transplant). Of the remaining 6 (46.2%) patients who engrafted and survived beyond 1 month of the transplant, the day 28 post-transplant MRD was negative for 5 patients while it was positive in 1 patient (0.13%). On follow up, 2 (15.4%) patients developed disease relapse (on days 54 and 218 respectively) and died. The remaining 4 (30.8%) patients are alive and relapse free at last follow up (mean follow up of surviving patients is 16 months). One patient received a CD34 cell boost on day 96 (cell dose - 8.55 x 10 6/kg) for poor graft function. For the entire cohort, the estimated 1-year event free survival is 28.8% ± 13.1%(Figure 1d). Whereas, acute GVHD was noted in 3 patients (50%; out of 6 evaluable patients) and chronic GVHD was noted in 3 patients (50%; out of the 6 evaluable patients). Thus, haploidentical NK cell therapy as an adjunct to transplant is safe and merits further evaluation in patients with AML. [Display omitted] DisclosuresMathews: Christian Medical College: Patents & Royalties: US 2020/0345770 A1 - Pub.Date Nov.5, 2020; AML: Other: Co-Inventor.

Complete Remission and Signs of Immunoediting Following Haploidentical NK Cell Therapy in Refractory High-Risk MDS and AML Ell Therapy in Refractory High-Risk MDS and AML

Haploidentical IL-2-activated natural killer (NK) cells can induce remission in patients with acute myeloid leukemia (AML). Here, we report results of a phase I/II clinical trial in which short-term IL-2-activated haploidentical NK cells were given to patients with primary chemotherapy-refractory or relapsed high-risk myelodysplastic syndrome (MDS), secondary AML (MDS/AML), and AML. The patients received conditioning with cyclophosphamide/fludarabine combined with total lymphoid irradiation. To avoid expansion of regulatory T cells (Treg), no post-NK cell infusion IL-2 was given. Using this protocol, nine of sixteen patients responded. Of the nine patients, six achieved a major response with complete remission (CR), marrow CR or partial response in combination with improved hematological parameters. Five of these patients had high-risk MDS or MDS/AML, and one had relapsed de novo AML. These patients all became eligible for, and proceeded to, allogeneic hematopoetic stem cell transplantation (HSCT). Four of the patients are still free from disease >4 years after treatment. The patients with a major responses had detectable donor NK cells at days 7 to 14 following infusion, displayed reduction of allelic burden, and partial or complete eradication of MDS/AML subclones. Residual lin-CD34+CD123+CD45RA+ blast cells showed an increase in total HLA class I- and HLA-E-expression suggesting immunoediting by the NK cells. Non/minor responders displayed an activation of CD4+ and CD8+ T cells as well as Treg cells accompanied by a marked increase in inflammatory cytokines. Together, this study supports the use of haploidentical NK cell infusions as a bridge to HSCT, especially for patients with high-risk MDS and MDS/AML. DisclosuresMiller:Celegene: Consultancy; Fate Therapeutics: Consultancy, Research Funding; Oxis Biotech: Consultancy. Hansson:Gilead: Honoraria, Research Funding; Abbvie: Honoraria; Janssen-Cilag: Honoraria, Research Funding.

Multi-Functional Genetic Engineering of Pluripotent Cell Lines for Universal Off-the-Shelf Natural Killer Cell Cancer Immunotherapy

Since the initial identification of natural killer (NK) cells, considerable effort has been made to harness their inherent anti-tumor capacities for the treatment of cancer. Despite recent clinical advancements utilizing the adoptive transfer of allogeneic NK cells, particularly with haplo-identical NK cells for acute myeloid leukemia, significant opportunities remain to further augment the anti-tumor activity of NK cells. Clinical trials have shown the persistence of adoptively transferred NK cells to be generally correlated with improved patient outcome. Natural killer cell persistence can be enhanced through several strategies, including by the administration of long-lived sub-populations such as adaptive-memory NK cells or of NK cells that are genetically modified to prolong survival or attenuate rejection by the host immune system. However, conventional sources of primary allogeneic NK cells have proven to be highly variable and difficult to genetically modify.Human induced pluripotent stem cells (hiPSCs) are unique in their capacity to self-renew and differentiate into any specialized cell type of the body. In addition, hiPSCs can be readily genetically-edited, and single hiPSCs can be clonally selected for the creation of engineered master pluripotent cell lines. As such, hiPSCs represent a renewable and reliable source for generating off-the-shelf engineered cell therapy products that are homogenous and display augmented anti-tumor properties.To enhance the persistence of hiPSC-derived NK cells, we employed a multi-dimensional strategy for hiPSC modification. First, hiPSCs were derived from fibroblast cells through a proprietary cellular reprogramming process that promotes telomere elongation. Second, hiPSCs were genetically-engineered to express cell surface-bound IL-15 to promote NK cell expansion during hiPSC differentiation and survival upon adoptive transfer. Third, additional hiPSC modifications were introduced to disrupt cell surface HLA Class I presentation and to drive cell surface expression of the non-classical HLA molecule HLA-G, to protect against host-mediated rejection by CD8+ T cells and NK cells, respectively. Lastly, we explored the influence of NKG2C to promote the adaptive-memory NK cell phenotype.We observed that hiPSC-derived NK cells displayed telomere length that was on average ~2-fold longer as compared to peripheral blood NK cells. The introduction of surface-bound IL-15 supported enhanced proliferation during differentiation with minimal requirements for cytokine support. Disruption of HLA-Class I expression decreased the proliferation of allogeneic CD8+ T cells upon challenge with modified hiPSCs as compared to wild-type hiPSCs (64% for wild-type vs. 29% for modified). This modification increased the cytotoxic activity of NK cells against modified hiPSCs (50% killing at 66 hours for modified as compared to 94 hours for wild-type). To this end, we further modified the hiPSCs to express the non-classical HLA molecule HLA-G. HLA-G protected against NK cell cytotoxicity and extended median survival of modified hiPSCs from 35 hours to 58 hours in an in vitro co-culture system. Finally, engineered hiPSC-derived NK cells can be effectively expanded (>1000x in 14 days) to support an efficient central manufacturing and banking method that enables multi-dose strategies to further support product persistence. Combined, the data suggest that our multi-engineered strategy can support the generation of a universal hiPSC backbone for the derivation of NK cells with improved persistence and anti-tumor efficacy. We are currently adding targeting modalities such as enhanced CD16 or chimeric antigen receptor expression to the discussed universal hiPSC backbone to generate master pluripotent cell lines to enable the manufacture of off-the-shelf NK cell cancer immunotherapy products. DisclosuresBjordahl:Fate Therapeutics: Employment, Equity Ownership. Clarke:Fate Therapeutics Inc.: Employment, Equity Ownership. Gaidarova:Fate Therapeutics: Employment. Bauer:Fate Therapeutics: Employment. Sasaki:Fate Therapeutics Inc.: Employment, Equity Ownership. Groff:Fate Therapeutics Inc.: Employment. Lee:Fate Therapeutics Inc.: Employment, Equity Ownership. Lan:Fate Therapeutics Inc.: Employment, Equity Ownership. Abujarour:Fate Therapeutics Inc.: Employment. Bonello:Fate Therapeutics Inc.: Employment. Robinson:Fate Therapeutics Inc.: Employment. Hardy:Fate Therapeutics: Employment. Valamehr:Fate Therapeutics: Employment, Equity Ownership.

Optimizing NK Cell-Based Immunotherapy in Myeloid Leukemia: Abrogating an Immunosuppressive Microenvironment.

Natural killer (NK) cells are prominent cytotoxic and cytokine-producing components of the innate immune system representing crucial effector cells in cancer immunotherapy. Presently, various NK cell-based immunotherapies have contributed to the substantial improvement in the reconstitution of NK cells against advanced-staged and high-risk AML. Various NK cell sources, including haploidentical NK cells, adaptive NK cells, umbilical cord blood NK cells, stem cell-derived NK cells, chimeric antigen receptor NK cells, cytokine-induced memory-like NK cells, and NK cell lines have been identified. Devising innovative approaches to improve the generation of therapeutic NK cells from the aforementioned sources is likely to enhance NK cell expansion and activation, stimulate ex vivo and in vivo persistence of NK cells and improve conventional treatment response of myeloid leukemia. The tumor-promoting properties of the tumor microenvironment and downmodulation of NK cellular metabolic activity in solid tumors and hematological malignancies constitute a significant impediment in enhancing the anti-tumor effects of NK cells. In this review, we discuss the current NK cell sources, highlight ongoing interventions in enhancing NK cell function, and outline novel strategies to circumvent immunosuppressive factors in the tumor microenvironment to improve the efficacy of NK cell-based immunotherapy and expand their future success in treating myeloid leukemia.

Cytokine-Induced Memory-Like NK Cells with High Reactivity against Acute Leukemia Blasts and Solid Tumor Cells Suitable for Adoptive Immunotherapy Approaches.

Simple SummarySeveral strategies have been under investigation on how to enhance anti-tumor responses in patients with poor prognosis. Considering the heterogeneity of antigens expressed by different hematological and solid neoplasia, it is difficult to develop specific immunotherapy approaches. The capacity of Natural Killer (NK) cells to kill tumor cells without specific Ag recognition provides an advantage over T cells and makes them appealing candidate effectors for immunotherapy, even if the effectiveness of their therapeutic use is limited by short-term persistence in vivo. After providing proof of concept regarding the feasibility of inducing NK cells endowed with anti-tumor activity and ability to persist for a protracted period in vivo, considering the relatively short period of in vitro activation, this approach could easily be translated into clinical practice to control tumor growth in high-risk patients.The limited efficacy of Natural Killer (NK) cell-based immunotherapy results in part from the suboptimal expansion and persistence of the infused cells. Recent reports suggest that the generation of NK cells with memory-like properties upon in vitro activation with defined cytokines might be an effective way of ensuring long-lasting NK cell function in vivo. Here, we demonstrate that activation with IL-12, IL-15 and IL-18 followed by a one-week culture with optimal doses of Interleukin (IL-2) and IL-15 generates substantial numbers of memory-like NK cells able to persist for at least three weeks when injected into NOD scid gamma (NSG) mice. This approach induces haploidentical donor-derived memory-like NK cells that are highly lytic against patients’ myeloid or lymphoid leukemia blasts, independent of the presence of alloreactive cell populations in the donor and with negligible reactivity against patients’ non-malignant cells. Memory-like NK cells able to lyse autologous tumor cells can also be generated from patients with solid malignancies. The anti-tumor activity of allogenic and autologous memory-like NK cells is significantly greater than that displayed by NK cells stimulated overnight with IL-2, supporting their potential therapeutic value both in patients affected by high-risk acute leukemia after haploidentical hematopoietic stem cell transplantation and in patients with advanced solid malignancies.

531 - BMT CTN 1803: Haploidentical Natural Killer Cells (K-NK002) to Prevent Post-Transplant Relapse in AML and MDS (NK-REALM)

Background and Rationale: Relapse remains the leading cause of treatment failure for patients with high-risk acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS) undergoing allogeneic bone marrow transplantation (BMT). Although the risk depends on parameters such as age and conditioning intensity, relapse is experienced by 30-50% of patients after conventional BMT in high-risk AML/MDS. Initial safety and post-BMT relapse risk reduction results were reported from a phase I study of haploidentical (haplo) donor-derived natural killer (NK) cells expanded ex vivo on feeder cells expressing IL-21 and 4-1BBL (FC21-NK) in conjunction with haploBMT. Of 13 patients with high-risk myeloid malignancies treated with FC21-NK cells, no infusion reactions or dose-limiting toxicities occurred and only 1 patient, treated at the lowest dose of 1×105 cells/kg, relapsed (Ciurea, Blood, 2017). This experience supports investigation of K-NK002, a product derived from haploidentical donor NK cells and expanded ex vivo in a feeder cell-free system using plasma membrane (PM21) particles bearing membrane-bound IL-21 and 4-1BBL. We used contemporary data from the Center for International Blood and Marrow Transplant Research registry to determine baseline relapse rates that informed the statistical design. K-NK002 will be given in the peri-transplant period to test the hypothesis that haploidentical NK cells can mediate an effective anti-leukemia response. Trial Design and Methods: BMT CTN 1803 is a phase II, single-arm, open-label, multicenter trial designed to investigate the safety and efficacy of K-NK002 for the treatment of patients with high-risk AML or MDS undergoing haploBMT (NCT04395092). An initial 6 patient safety run-in phase will precede enrollment into the full study of approximately 60 patients. Major inclusion criteria of patients and donors are listed in Table 1. Production of the haploidentical donor NK-cells is completed prior to the planned haploBMT. BMT conditioning will consist of 140 mg/m2 (100 mg/m2 for patients ≥60 years old) melphalan on Day -7; 40 mg/m2 fludarabine on Days -7, -6, -5, and -4; and 2 Gy of total body irradiation on Day -3. Donor bone marrow will be harvested and given on Day 0. Three doses of K-NK002 (1×108 NK cells/kg) will be administered IV on Days -2, +7, and +28, relative to the haploBMT. Graft-versus-host disease (GVHD) prophylaxis consists of post-transplantation cyclophosphamide with tacrolimus and mycophenolate mofetil. The primary endpoint is cumulative incidence of relapse at 1 year post haploBMT in patients receiving at least 1 infusion of K-NK002. Secondary endpoints are safety and tolerability of K-NK002; overall survival; non-relapse mortality; relapse-free survival; GVHD-free survival; cumulative incidence of acute GVHD and chronic GVHD; hematologic recovery; donor-cell engraftment; primary and secondary graft failure; overall incidence of toxicity; and cumulative incidence of infections including cytomegalovirus re-activation and symptomatic BK virus hemorrhagic cystitis. Exploratory endpoints are systemic immunosuppression-free survival; immune reconstitution at Days 28, 100, and 365 post haploBMT; proportion of patients with detectable minimal residual disease at Days 28 and 100 post haploBMT; feasibility of administering the planned K-NK002 doses; and impact of NK-cell alloreactivity on relapse and survival. Download : Download high-res image (319KB) Download : Download full-size image Disclosures Bejanyan: Kiadis Pharma: Membership on an entity’s Board of Directors or advisory committees. Devine: Magenta Therapeutics: Consultancy. Luznik: WindMil Therapeutics: Patents & Royalties: Patent holder; Genentech: Research Funding; Merck: Research Funding, Speakers Bureau; AbbVie: Consultancy. Sandler: Kiadis Pharma: Current Employment. Krakow: HighPass Bio: Research Funding. Fitzgerald: Kiadis Pharma: Current Employment. Tracey: Kiadis Pharma: Current Employment. Champlin: DKMS America: Membership on an entity’s Board of Directors or advisory committees; Takeda: Patents & Royalties; Actinium: Consultancy; Johnson and Johnson: Consultancy; Cytonus: Consultancy; Omeros: Consultancy; Genzyme: Speakers Bureau.

Cellular immunotherapy with multiple infusions of in vitro-expanded haploidentical natural killer cells after autologous transplantation for patients with plasma cell myeloma

Background aimsTo investigate the feasibility and safety of haploidentical natural killer (NK) cell infusions as consolidation immunotherapy after autologous stem cell transplant (ASCT) in patients with plasma cell myeloma. MethodsTen patients (median age, 59 years) received induction treatment followed by high-dose melphalan (200 mg/m2) at day –1, ASCT at day 0 and increasing NK cell doses (1.5 × 106, 1.5 × 107 and multiple doses of 1.0 × 108 cells/kg body weight) from day +1 to day +30 after ASCT. NK cells were harvested and purified from peripheral blood of haploidentical donors and expanded for 19 days with interleukin (IL)-2 and IL-15 under Good Manufacturing Practice conditions. ResultsNK cell numbers increased 56.0-fold (37.4- to 75.5-fold). Patients received a median of 3.8 × 108 (0.9–5.7 × 108) NK cells/kg body weight in six (three to eight) infusions. Multiparametric mass cytometry analysis demonstrated an altered surface receptor repertoire of expanded NK cells with increased degranulation and cytokine production activities but diminished expression of perforin. Donor NK cells were detectable in the peripheral blood, peaking 1 h after each dose (up to 90% donor NK cells). The treatment was safe and well tolerated, without evidence of graft-versus-host disease. Comparison with a control patient population receiving ASCT without NK cell infusions showed no significant difference in relapse, progression-free survival and overall survival. ConclusionsThis study demonstrates reliable manufacturing of high numbers of activated NK cells for multiple-dose infusions and safe administration of these cellular products. The trial was registered at ClinicalTrials.gov (identifier no. NCT01040026).

BMT CTN 1803: Haploidentical Natural Killer Cells (K-NK002) to Prevent Post-Transplant Relapse in AML and MDS (NK-REALM)

Background and Rationale: Relapse remains the leading cause of treatment failure for patients with high-risk acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS) undergoing allogeneic bone marrow transplantation (BMT). Although the risk depends on parameters such as age and conditioning intensity, relapse is experienced by 30-50% of patients after conventional BMT in high-risk AML/MDS. Initial safety and post-BMT relapse risk reduction results were reported from a phase I study of haploidentical (haplo) donor-derived natural killer (NK) cells expanded ex vivo on feeder cells expressing IL-21 and 4-1BBL (FC21-NK) in conjunction with haploBMT. Of 13 patients with high-risk myeloid malignancies treated with FC21-NK cells, no infusion reactions or dose-limiting toxicities occurred and only 1 patient, treated at the lowest dose of 1×105 cells/kg, relapsed (Ciurea, Blood, 2017). This experience supports investigation of K-NK002, a product derived from haploidentical donor NK cells and expanded ex vivo in a feeder cell-free system using plasma membrane (PM21) particles bearing membrane-bound IL-21 and 4-1BBL. We used contemporary data from the Center for International Blood and Marrow Transplant Research registry to determine baseline relapse rates that informed the statistical design. K-NK002 will be given in the peri-transplant period to test the hypothesis that haploidentical NK cells can mediate an effective anti-leukemia response. Trial Design and Methods: BMT CTN 1803 is a phase II, single-arm, open-label, multicenter trial designed to investigate the safety and efficacy of K-NK002 for the treatment of patients with high-risk AML or MDS undergoing haploBMT (NCT04395092). An initial 6 patient safety run-in phase will precede enrollment into the full study of approximately 60 patients. Major inclusion criteria of patients and donors are listed in Table 1. Production of the haploidentical donor NK-cells is completed prior to the planned haploBMT. BMT conditioning will consist of 140 mg/m2 (100 mg/m2 for patients ≥60 years old) melphalan on Day -7; 40 mg/m2 fludarabine on Days -7, -6, -5, and -4; and 2 Gy of total body irradiation on Day -3. Donor bone marrow will be harvested and given on Day 0. Three doses of K-NK002 (1×108 NK cells/kg) will be administered IV on Days -2, +7, and +28, relative to the haploBMT. Graft-versus-host disease (GVHD) prophylaxis consists of post-transplantation cyclophosphamide with tacrolimus and mycophenolate mofetil. The primary endpoint is cumulative incidence of relapse at 1 year post haploBMT in patients receiving at least 1 infusion of K-NK002. Secondary endpoints are safety and tolerability of K-NK002; overall survival; non-relapse mortality; relapse-free survival; GVHD-free survival; cumulative incidence of acute GVHD and chronic GVHD; hematologic recovery; donor-cell engraftment; primary and secondary graft failure; overall incidence of toxicity; and cumulative incidence of infections including cytomegalovirus re-activation and symptomatic BK virus hemorrhagic cystitis. Exploratory endpoints are systemic immunosuppression-free survival; immune reconstitution at Days 28, 100, and 365 post haploBMT; proportion of patients with detectable minimal residual disease at Days 28 and 100 post haploBMT; feasibility of administering the planned K-NK002 doses; and impact of NK-cell alloreactivity on relapse and survival. Disclosures Bejanyan: Kiadis Pharma: Membership on an entity's Board of Directors or advisory committees. Devine:Magenta Therapeutics: Consultancy. Luznik:WindMil Therapeutics: Patents &amp; Royalties: Patent holder; Genentech: Research Funding; Merck: Research Funding, Speakers Bureau; AbbVie: Consultancy. Sandler:Kiadis Pharma: Current Employment. Krakow:HighPass Bio: Research Funding. Fitzgerald:Kiadis Pharma: Current Employment. Tracey:Kiadis Pharma: Current Employment. Champlin:DKMS America: Membership on an entity's Board of Directors or advisory committees; Takeda: Patents &amp; Royalties; Actinium: Consultancy; Johnson and Johnson: Consultancy; Cytonus: Consultancy; Omeros: Consultancy; Genzyme: Speakers Bureau.

Effect of Recombinant Human Adenovirus Type 5 Injection Combined with Haploidentical Natural Killer Cell Immunotherapy on Immune Function and Adverse Reactions in Patients with Malignant Ascites Caused by Ovarian Cancer

Objective: To explore the effect of recombinant human adenovirus type 5 injection combined with haploidentical natural killer cell immunotherapy on immune function and adverse reactions in patients with malignant ascites caused by ovarian cancer (OC). Methods: From January 2018 to September 2019, 65 patients with advanced OC combined with malignant ascites who received treatment in the oncology department of our hospital were collected. According to the treatment methods, the patients were assigned to study group (SG) (35 cases) and control group (CG) (30 cases). In CG, patients were treated with NK cell reinfusion. SG: On the basis of the CG, the recombinant human adenovirus type 5 injection was injected directly into tumour. The peripheral blood mononuclear cells of lineal relatives of all patients were cultivated by antibody coating and co-stimulation with various cytokines, and the NK cells cultured in vitro were returned to the patients for infusion twice on the 14th and 15th days, respectively. The clinical efficacy, changes of serum tumour markers, changes of immune function indicators and adverse effects were compared between the two groups. Results: The objective remission rate of clinical efficacy in the SG was obviously higher than that in the CG. Before therapy, the levels of serum carcinoembryonic antigen, carbohydrate antigen 242, carbohydrate antigen 19-9, alpha-fetoprotein and neuron specific enolase had no statistically significant difference in both groups (P&gt;0.05). After therapy, the level of each index in the SG was better than that in the CG, with statistically significant difference (P&lt;0.05). After therapy, the levels of peripheral NK cells, CD4+/CD8+ ratio, CD3+CD4+ and CD3+ level in SG were obviously higher than those before treatment, while the levels of CD4+CD25+ were obviously lower than those before treatment, and the indexes in the SG were obviously better than those in the CG (P&lt;0.05). Before therapy, there was no obvious difference in quality of life between the two groups. After treatment, the overall quality of life in the SG was higher than that in the CG, with statistically significant difference (P&lt;0.05). The incidence of hepatic and renal insufficiency had no statistically significant difference between SG and CG (P&gt;0.05). However, the incidence of nausea, vomiting, abdominal pain and diarrhea in the SG was obviously lower than that in the CG, with statistically significant difference (P&lt;0.05). Conclusion: Recombinant human adenovirus type 5 injection combined with haploidentical natural killer cell immunotherapy can better recover the immune function and reduce the incidence of adverse reactions in patients with malignant ascites caused by OC.

Exploring the NK cell platform for cancer immunotherapy.

Natural killer (NK) cells are cytotoxic lymphocytes of the innate immune system that are capable of killing virally infected and/or cancerous cells. Nearly 20 years ago, NK cell-mediated immunotherapy emerged as a safe and effective treatment approach for patients with advanced-stage leukaemia. Subsequently, the field of NK cell-based cancer therapy has grown exponentially and currently constitutes a major area of immunotherapy innovation. In general, the development of NK cell-directed therapies has two main focal points: optimizing the source of therapeutic NK cells for adoptive transfer and enhancing NK cell cytotoxicity and persistence in vivo. A wide variety of sources of therapeutic NK cells are currently being tested clinically, including haploidentical NK cells, umbilical cord blood NK cells, stem cell-derived NK cells, NK cell lines, adaptive NK cells, cytokine-induced memory-like NK cells and chimeric antigen receptor NK cells. A plethora of methods to augment the cytotoxicity and longevity of NK cells are also under clinical investigation, including cytokine-based agents, NK cell-engager molecules and immune-checkpoint inhibitors. In this Review, we highlight the variety of ways in which diverse NK cell products and their auxiliary therapeutics are being leveraged to target human cancers. We also identify future avenues for NK cell therapy research.

A phase I clinical trial testing the safety of IL-21-expanded, off-the-shelf, natural killer cells for relapsed/refractory acute myeloid leukemia and myelodysplastic syndrome.

TPS7562 Background: Allogeneic transplantation (Allo-HCT) demonstrates the enduring and potent role of the immune system in the control and eradication of acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). However, patients with relapsed, refractory (R/R) disease or comorbidities are not eligible for Allo-HCT. We sought to develop an allogeneic Natural killer (NK) cell-based immunotherapy approach to induce remission for these patients. The efficacy of haploidentical NK cells expanded ex vivo using a K562 feeder-cell line transfected with IL-21 and 41BBL has been established in R/R AML patients. However, haploidentical donor-derived NK cell manufacturing exceeds three weeks with the possibility of fulminant malignancy rendering patients ineligible for cellular therapy. To address this limitation we established a third-party NK cell bank derived from KIR and HLA-mismatched ‘ideal’ donors that allows scalable, affordable mass-production of large numbers of NK cells suitable for banking and immediate ‘off-the-shelf’ (OTS) administration to a broad population of recipients. Methods: This phase I study follows a 3+3 design to investigate the safety of mIL-21-expanded, third-party, OTS NK cells for treatment of R/R AML and MDS patients. Patients aged ≥18 or ≤80 years are enrolled into two cohorts: those &lt;60 years and able to tolerate intensive chemo will receive Fludarabine 30mg/m2/day (days -6 to -2) and Cytarabine 2g/m2/day (days -6 to -2). Patients &gt;60 years or &lt;60 years and unable/unwilling to tolerate intensive chemo will receive Fludarabine 30mg/m2/day (days -5 to -2) and Decitabine 20mg/m2/day (days -6 to -2). All patients subsequently receive a total of 6 infusions of NK cells administered thrice weekly for two weeks (between days 0-21) and will be followed up to day 56 from first NK cell infusion. Three NK cell dose-levels: 1x107, 3x107 and 1x108 cells/kg/dose will be explored to determine maximum tolerated dose (MTD). 3-18 patients/cohort/dose may be enrolled for MTD determination plus an additional 10 patients/dose in an expansion phase (maximum 28/cohort = 56 total subjects). Primary objective is to determine safety and feasibility of NK cell infusions. Secondary objectives will explore rates of remission PFS, overall survival and measurable residual disease negativity, cell counts, infectious complications, and patients proceeding to transplant. Enrollment in dose level 1 has started. Clinical trial information: NCT04220684 .