D - Statement of Work.pdf

Version 11-30-2016-1 FAR Part 11—Describing Agency Needs

STATEMENT OF WORK (SERVICES)

(SOW)

GENERAL INFORMATION

Title of Project:

snRNAseq analysis of senolytics in the aging kidney and lung

Statement of Need and Purpose:

The purpose of this contract is to procure RNA sequencing and data analysis services (following tissue dissociation, nuclei extraction, library preparation) from precious mouse kidney and lung tissues dissected from senolytic treated mice at different time points to identify transcriptomic changes with age and senolytic treatment of these tissues.

Background Information and Objective:

Single-cell RNA sequencing (scRNAseq) emerges as an indispensable technique for dissecting the dynamics of senescence induction following drug administration in mice, particularly in organs as diverse as the kidney and lung. The complexity of these tissues requires a cellular-level investigation to unravel the intricacies of senescence dynamics. By employing scRNAseq, The National Institute on Aging (NIA) can delve into the heterogeneous cellular responses, uncovering nuanced changes in gene expression profiles across various cell types over time.

This approach not only provides a comprehensive understanding of senescence progression but also unveils potential cell-specific therapeutic targets, advancing the ability to intervene in age-related pathologies.

Aging is a multifaceted process that affects all organisms, and understanding its mechanisms is crucial for developing interventions to promote healthy aging. The kidney, a vital organ responsible for filtration and waste excretion, undergoes significant changes with age, leading to functional decline and increased susceptibility to disease. Investigating the dynamics of senescence within the kidney is essential for elucidating the molecular pathways involved in renal aging.

Among vital organs, the lungs play a pivotal role in oxygen exchange, facing significant alterations with age, which lead to functional deterioration and heightened disease susceptibility.

Exploring the dynamics of senescence within the pulmonary tissue is imperative for unraveling the molecular pathways implicated in pulmonary aging.

This project aims to study senescence dynamics in the kidney and lungs of mice by administering doxorubicin, a chemotherapeutic agent. Following doxorubicin administration, mice will be sacrificed at various time points (1, 2, 4, 6, and 12 days), and their kidneys and lungs will be harvested for analysis. NIA will employ a technique known as single nuclei RNA sequencing (snRNA-seq) to examine the transcriptomic landscape of renal cells and pulmonary cells at each time point.

These organs are complex and are composed of diverse cell populations, each playing unique roles in maintaining function. Traditional RNA sequencing methods often fail to capture this cellular heterogeneity, leading to a limited understanding of gene expression changes occurring during senescence. snRNA-seq overcomes this limitation by enabling transcriptomic analysis at the single-cell level. By leveraging snRNA-seq, NIA can dissect the distinct transcriptional responses of individual cell types to senescence-inducing stimuli. This information will provide valuable insights into the functional consequences of senescence on different cell populations and identify potential therapeutic targets to mitigate age-related organ dysfunction.

Period of Performance:

Within nine (9) months after the date of award.

SCOPE OF WORK

General Requirements:

Independently and not as an agent of the Government, the Contractor shall furnish all the necessary services, qualified personnel, material, equipment, and facilities, not otherwise provided by the Government as needed to perform the Statement of Work below:

Specific Requirements:

NIA requires snRNAseq. NIA has already collected and frozen all the samples. The only way to perform single-cell RNAseq in frozen tissue pieces is through the technique proposed here, snRNAseq. The services required for this project include:

• Nuclei isolation for 36 samples (18 lung samples and 18 kidney samples)

• RNA extraction from the nuclei

• Single-nuclei RNA library preparation using 10X Genomics platform

• Illumina Nova Seq (250M reads/sample)

LEVEL OF EFFORT: N/A

GOVERNMENT RESPONSIBILITIES

The government will provide the samples and is responsible for reviewing and approving final reports generated under the contract.

DELIVERY OR DELIVERABLES

The vendor shall process the samples and provide sequencing data and analysis within 3 months of receipt of the samples. The vendor shall provide progress updates during the entire process. The data and analysis shall be provided electronically.

REPORTING REQUIREMENTS

The vendor shall provide the sequencing data and analysis electronically within 3 months of receipt of the samples.

OTHER CONSIDERATIONS

Travel:

No travel reimbursement is authorized for this requirement.

Information System Security Plan:

The vendor does not require access to a federal information system(s).

Data Rights:

The data produced by this project will belong to the government to be used in further data analysis and publications. The government will have unlimited rights.