Testicular vein size as measured by scrotal ultrasonography is independently predictive of varicocelectomy outcomes for men with clinical grade 1 and 2 varicoceles

Abstract Background Clinical-grade whole-genome sequencing (cWGS) has the potential to become the standard of care within the clinic because of its breadth of coverage and lack of bias towards certain regions of the genome. Colorectal cancer presents a difficult treatment paradigm, with over 40% of patients presenting at diagnosis with metastatic disease. We hypothesised that cWGS coupled with 3′ transcriptome analysis would give new insights into colorectal cancer. Methods Patients underwent PCR-free whole-genome sequencing and alignment and variant calling using a standardised pipeline to output SNVs, indels, SVs and CNAs. Additional insights into the mutational signatures and tumour biology were gained by the use of 3′ RNA-seq. Results Fifty-four patients were studied in total. Driver analysis identified the Wnt pathway gene APC as the only consistently mutated driver in colorectal cancer. Alterations in the PI3K/mTOR pathways were seen as previously observed in CRC. Multiple private CNAs, SVs and gene fusions were unique to individual tumours. Approximately 30% of patients had a tumour mutational burden of > 10 mutations/Mb of DNA, suggesting suitability for immunotherapy. Conclusions Clinical whole-genome sequencing offers a potential avenue for the identification of private genomic variation that may confer sensitivity to targeted agents and offer patients new options for targeted therapies.

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Inference of molecular structure for characterization and improvement of clinical grade immunocytokines

Journal of Structural Biology ◽

10.1016/j.jsb.2021.107696 ◽

2021 ◽

Vol 213 (1) ◽

pp. 107696

Author(s):

Tiziano Ongaro ◽

Salvatore R. Guarino ◽

Luigi Scietti ◽

Martina Palamini ◽

Sarah Wulhfard ◽

...

Keyword(s):

Molecular Structure ◽

Clinical Grade

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Optimizing the Procedure to Manufacture Clinical-Grade NK Cells for Adoptive Immunotherapy

Cancers ◽

10.3390/cancers13030577 ◽

2021 ◽

Vol 13 (3) ◽

pp. 577

Author(s):

Adrián Fernández ◽

Alfonso Navarro-Zapata ◽

Adela Escudero ◽

Nerea Matamala ◽

Beatriz Ruz-Caracuel ◽

...

Keyword(s):

Nk Cells ◽

Cell Activation ◽

Mononuclear Cells ◽

Nk Cell ◽

Fold Increase ◽

Clinical Grade ◽

Transcriptome Profile ◽

Expansion Process ◽

Peripheral Blood Mononuclear ◽

Activation Status

Natural killer (NK) cells represent promising tools for cancer immunotherapy. We report the optimization of an NK cell activation–expansion process and its validation on clinical-scale. Methods: RPMI-1640, stem cell growth medium (SCGM), NK MACS and TexMACS were used as culture mediums. Activated and expanded NK cells (NKAE) were obtained by coculturing total peripheral blood mononuclear cells (PBMC) or CD45RA+ cells with irradiated K562mbIL15-41BBL or K562mbIL21-41BBL. Fold increase, NK cell purity, activation status, cytotoxicity and transcriptome profile were analyzed. Clinical-grade NKAE cells were manufactured in CliniMACS Prodigy. Results: NK MACS and TexMACs achieved the highest NK cell purity and lowest T cell contamination. Obtaining NKAE cells from CD45RA+ cells was feasible although PBMC yielded higher total cell numbers and NK cell purity than CD45RA+ cells. The highest fold expansion and NK purity were achieved by using PBMC and K562mbIL21-41BBL cells. However, no differences in activation and cytotoxicity were found when using either NK cell source or activating cell line. Transcriptome profile showed to be different between basal NK cells and NKAE cells expanded with K562mbIL21-41BBL or K562mbIL15-41BBL. Clinical-grade manufactured NKAE cells complied with the specifications from the Spanish Regulatory Agency. Conclusions: GMP-grade NK cells for clinical use can be obtained by using different starting cells and aAPC.

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