2203

OBJECTIVES/SPECIFIC AIMS: To date, only 1 documented case of an individual cured of HIV has been reported. He received an allogeneic bone marrow transplant with cells harboring an HIV-resistant genotype. To mimic this result, we have initiated a Phase I to evaluate the safety of an autologous stem cell gene therapy bone marrow transplant in HIV-related lymphoma patients. METHODS/STUDY POPULATION: The first cohort of patients will receive a 1:1 ratio of unmanipulated CD34 hematopoietic stem cells (HSC) and lentivector modified CD34 HSC expressing a combination of HIV-resistant genes and a selectable marker for cell sorting prior to transplantation. Safety of the HIV-resistant stem cells will be assessed by evaluating engraftment, expression of the anti-HIV genes, and the stability and sequence of the vector. RESULTS/ANTICIPATED RESULTS: One patient has been enrolled and transplanted with the HIV-resistant stem cells. After 1 and 2 months post-transplant, patient blood samples were received, processed for genomic DNA, analyzed by quantitative PCR (qPCR), and displayed successful engraftment of 16 and 12 vector copies per 100 cells, respectively. Expression of all anti-HIV genes was confirmed by qPCR. PCR on genomic DNA confirmed the correct sizes and sequences of the integrated vector and confirmed the successful engraftment of our gene modified cells. Currently, we are enrolling more patients into the trial. DISCUSSION/SIGNIFICANCE OF IMPACT: If successful, this therapy has the potential to change HIV treatment.

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In Vivo Selection of Antifolate-Resistant Transgenic Hematopoietic Stem Cells in a Murine Bone Marrow Transplant Model

Journal of Pharmacology and Experimental Therapeutics ◽

10.1124/jpet.300.1.50 ◽

2002 ◽

Vol 300 (1) ◽

pp. 50-56 ◽

Cited By ~ 15

Author(s):

Christopher A. Warlick ◽

Michaeleen D. Diers ◽

John E. Wagner ◽

R. Scott McIvor

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Bone Marrow Transplant ◽

Marrow Transplant ◽

Hematopoietic Stem ◽

Murine Bone Marrow ◽

In Vivo Selection ◽

Transplant Model ◽

Selection Of

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Impact of Treatment Prior to Allogeneic Transplantation of Hematopoietic Stem Cells in Patients with Myelodysplastic Syndrome: Results of the Latin American Bone Marrow Transplant Registry

Biology of Blood and Marrow Transplantation ◽

10.1016/j.bbmt.2020.01.030 ◽

2020 ◽

Vol 26 (5) ◽

pp. 1021-1024

Author(s):

Fernando Barroso Duarte ◽

Anna Thawanny Gadelha Moura ◽

Vaneuza Araújo Moreira Funke ◽

Virgílio Antônio Rensi Colturato ◽

Nelson Hamerschlak ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Myelodysplastic Syndrome ◽

Hematopoietic Stem Cells ◽

Bone Marrow Transplant ◽

Latin American ◽

Allogeneic Transplantation ◽

Marrow Transplant ◽

Hematopoietic Stem ◽

Transplant Registry

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Multimodal Tracking of Hematopoietic Stem Cells from Young and Old Mice Labeled with Magnetic–Fluorescent Nanoparticles and Their Grafting by Bioluminescence in a Bone Marrow Transplant Model

Biomedicines ◽

10.3390/biomedicines9070752 ◽

2021 ◽

Vol 9 (7) ◽

pp. 752

Author(s):

Fernando A. Oliveira ◽

Mariana P. Nucci ◽

Javier B. Mamani ◽

Arielly H. Alves ◽

Gabriel N. A. Rego ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Hematopoietic Stem Cells ◽

Bone Marrow Transplant ◽

Marrow Transplant ◽

Hematopoietic Stem ◽

Icp Ms ◽

Old Mice

This study proposes an innovative way to evaluate the homing and tracking of hematopoietic stem cells from young and old mice labeled with SPIONNIRF-Rh conjugated with two types of fluorophores (NIRF and Rhodamine), and their grafting by bioluminescence (BLI) in a bone marrow transplant (BMT) model. In an in vitro study, we isolated bone marrow mononuclear cells (BM-MNC) from young and old mice, and analyzed the physical–chemical characteristics of SPIONNIRF-Rh, their internalization, cell viability, and the iron quantification by NIRF, ICP-MS, and MRI. The in vivo study was performed in a BMT model to evaluate the homing, tracking, and grafting of young and old BM-MNC labeled with SPIONNIRF-Rh by NIRF and BLI, as well as the hematological reconstitution for 120 days. 5FU influenced the number of cells isolated mainly in young cells. SPIONNIRF-Rh had adequate characteristics for efficient internalization into BM-MNC. The iron load quantification by NIRF, ICP-MS, and MRI was in the order of 104 SPIONNIRF-Rh/BM-MNC. In the in vivo study, the acute NIRF evaluation showed higher signal intensity in the spinal cord and abdominal region, and the BLI evaluation allowed follow-up (11–120 days), achieving a peak of intensity at 30 days, which remained stable around 108 photons/s until the end. The hematologic evaluation showed similar behavior until 30 days and the histological results confirm that iron is present in almost all tissue evaluated. Our results on BM-MNC homing and tracking in the BMT model did not show a difference in migration or grafting of cells from young or old mice, with the hemogram analysis trending to differentiation towards the myeloid lineage in mice that received cells from old animals. The cell homing by NIRF and long term cell follow-up by BLI highlighted the relevance of the multimodal nanoparticles and combined techniques for evaluation.

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