Engineering NK-CAR.19 cells with the IL-15/IL-15Rα complex improved proliferation and anti-tumor effect in vivo

IntroductionNatural killer 92 (NK-92) cells are an attractive therapeutic approach as alternative chimeric antigen receptor (CAR) carriers, different from T cells, once they can be used in the allogeneic setting. The modest in vivo outcomes observed with NK-92 cells continue to present hurdles in successfully translating NK-92 cell therapies into clinical applications. Adoptive transfer of CAR-NK-92 cells holds out the promise of therapeutic benefit at a lower rate of adverse events due to the absence of GvHD and cytokine release syndrome. However, it has not achieved breakthrough clinical results yet, and further improvement of CAR-NK-92 cells is necessary.MethodsIn this study, we conducted a comparative analysis between CD19-targeted CAR (CAR.19) co-expressing IL-15 (CAR.19-IL15) with IL-15/IL-15Rα (CAR.19-IL15/IL15Rα) to promote NK cell proliferation, activation, and cytotoxic activity against B-cell leukemia. CAR constructs were cloned into lentiviral vector and transduced into NK-92 cell line. Potency of CAR-NK cells were assessed against CD19-expressing cell lines NALM-6 or Raji in vitro and in vivo in a murine model. Tumor burden was measured by bioluminescence.ResultsWe demonstrated that a fourth- generation CD19-targeted CAR (CAR.19) co-expressing IL-15 linked to its receptor IL-15/IL-15Rα (CAR.19-IL-15/IL-15Rα) significantly enhanced NK-92 cell proliferation, proinflammatory cytokine secretion, and cytotoxic activity against B-cell cancer cell lines in vitro and in a xenograft mouse model.ConclusionTogether with the results of the systematic analysis of the transcriptome of activated NK-92 CAR variants, this supports the notion that IL-15/IL-15Rα comprising fourth-generation CARs may overcome the limitations of NK-92 cell-based targeted tumor therapies in vivo by providing the necessary growth and activation signals

Lower insulin-dose adjusted A1c (IDAA1c) is associated with less complications in Individuals with Type 1 Diabetes treated with hematopoetic stem-cell transplantation and conventional therapy

Objective: To evaluate the association between insulin-dose adjusted A1C (IDAA1c) and microvascular complications (MC) and hypoglycemia in a representative Brazilian population of Type 1 diabetes mellitus (T1DM) patients. Research Design and Methods: This was a cross-sectional study based on a previous study, “Microvascular Complications in Type 1 Diabetes: a comparative analysis of patients treated with autologous nonmyeloablative hematopoietic stem-cell transplantation (AHST) and conventional medical therapy (CT)”. The 168 patients in that study (144 from CT plus 24 from AHST) were re-subdivided into two groups, according to their IDAA1c values (30 patients had IDAA1c ≤ 9; 138 had IDAA1c > 9). Then, the prevalence of MC (diabetic renal disease, neuropathy, and retinopathy), hypoglycemia (blood glucose <60 mg/dL), and severe hypoglycemic (episode of hypoglycemia that required the assistance of another person to treat) events were compared between the groups. The groups were well-matched on these factors: duration of disease, sex, and age at the time of diagnosis of T1DM. Results: After an average of 8 years after diagnosis, only 6.6% (2/30) of the patients from IDAA1c ≤ 9 group developed any MC, whereas 21.0% (29/138) from the IDAA1c > 9 group had at least one complication (p = 0.044). Regarding hypoglycemic events, the proportion of individuals who reported at least 1 episode of hypoglycemia in the last month was 43.3 and 64.7% from the IDAA1c ≤ 9 and IDAA1c > 9 groups, respectively (p = 0.030). Regarding severe hypoglycemia, the proportion of patients presenting at least one episode in the last month and the rate of episode/patient/month were similar between groups (6.7 vs. 13.2%; p = 0.535; and 0.1/patient/month vs. 0.25/patient/month; p = 0.321). Conclusion: In a representative Brazilian population of T1DM patients, those with IDAA1c ≤ 9 presented a lower frequency of MC, as well as fewer episodes of hypoglycemia, in the month prior to the analysis.publishedVersio

Emerging CAR T cell therapies: clinical landscape and patent technological routes

The purpose of this study is to mine CAR-T patents and therapies under development, to design a landscape of the sector and to understand key therapy segments and their current trends. The study analyzed the entire market, consisting of 1624 patent families and 509 biologics under development, to depict an overview of the CAR-T therapies and their state of the art. Our results showed cutting-edge inventions, the major players, the dynamics of cooperation among institutions, the progress of the therapies’ generation over the years and future innovation pathways. CAR-T therapies are transforming the current scenario for cancer treatment, and this study reveals the picture of what we can likely expect ahead in order to assist scientists at the academy and industry to improve their research strategies

Bone marrow-derived MSCs isolated from <i>Wistar</i> rats.

<p>Immunologic phenotypes of MSCs (A). Passage five plastic adherent cells cultured in alpha-MEM supplemented with 15% fetal bovine serum (B) and differentiated into adipogenic (C) or osteogenic (D) lineages.</p

Glucose Tolerance Test (GTT) (A) and Insulin Tolerance Test (ITT) glycemia responses (B) increase in high fat diet-fed mice.

<p>Glycemia AUC, area under the curve (AUC) analysis of glycemia profile of GTT and ITT after eight weeks of diet regimen. Values are expressed as mean ± SEM (10–28 mice/group). * p < 0.05, control group (C) <i>vs</i>. high fat diet group (HFD).</p

Islet apoptosis increases in HFD-PBS and HFD-MSCs-NR and decreases in HFD-MSCs-R.

<p>Representative images of pancreas section stained for caspase-3 of control group (A) and HFD-PBS (B), HFD-MSCs-R (C), and HFD-MSCs-NR (D) groups. Quantitative data correspond to mean ± SEM (5–13 mice/group) (E). * p < 0.05, control group (C) <i>vs</i>. HFD-PBS; ^# p < 0.05, HFD-MSCs-R <i>vs</i>. HFD-PBS; ⁺ p < 0.05, HFD-MSCs-NR <i>vs</i>. HFD-PBS; ^& p < 0.05, HFD-MSCs-NR <i>vs</i>. HFD-MSCs-R.</p

Glucose Tolerance Test (GTT) glycemia response decrease post-MSCs infusion.

<p>Only in the 16^th week post-MSC infusions, the glycemia response to glucose injection was significantly lower in HFD-MSCs-R when compared to HFD-PBS group. Values are expressed as mean ± SEM (5–13 mice/group). * p < 0.05, control group (C) <i>vs</i>. HFD-PBS; ^# p < 0.05, HFD-MSCs-R <i>vs</i>. HFD-PBS.</p